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2024-03-28T15:30:06Z
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http://2014.igem.org/Team:UMaryland/project/biobricks
Team:UMaryland/project/biobricks
2014-10-17T20:40:49Z
<p>Nbb826: </p>
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<h1 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/project/biobricks " class="contentpagetitle"> <br />
Biobricks <br />
</a><br />
</h1><br />
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<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; text-decoration: underline;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">Part:BBa_K1489002 | </span></span><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap; text-decoration: underline;">Lpp-OmpA in new backbone</span></strong></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/parts/f/fd/PSB1C3_-_BBa_K1489002.png" border="0" width="480" height="335" style="border: 0;" /></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;">We moved an existing biobrick, BBa_K103006, from an old plasmid backbone, pSB1A2, into the current backbone, pSB1C3, via RE cloning.</span></p><br />
<p style="text-align: center;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 20px; font-weight: bold; line-height: 19.5px; text-align: start;">Lpp-OmpA-Linker (pSB1C3)</span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">Outer membrane protein A (OmpA) is a common outer membrane protein found in bacteria that serves various purposes. For our purposes, the beta-barrel transmembrane domain can be utilized to anchor otherwise soluble proteins in the outer cell membrane. Lpp serves to direct the fusion protein, while a unstructured linker (GGGSGGGS) serves to separate OmpA from its cargo in order to avoid interactions between the two proteins.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">The expressed protein is structurally identical to BBa_K103006. This version of the biobrick has been moved into the new backbone, pSB1C3, which contains the gene for chloramphenicol resistance. This resistance gene contains an internal SacI site, making the SacI site at the end of the gene almost unusable for RE cloning. BBa_K1489003 mutagenizes this SacI site into a KasI in order to avoid this issue.</p><br />
<p><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;"> </span></p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="line-height: 12.0000009536743px; color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; text-decoration: underline;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">Part:BBa_K1489000 | </span></span><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap; text-decoration: underline;">pBAD-RBS-Bovine Galectin 1-B0012 Terminator</span></strong></span></p><br />
<p><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/parts/4/42/PSB1C3_-_BBa_K1489000.png" border="0" width="515" height="335" style="border: 0px; display: block; margin-left: auto; margin-right: auto;" /></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;">A soluble his-tagged galectin from Bos taurus, expressed under Arabinose induction</span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px; text-align: center;"><span style="font-size: 20px; font-weight: bold;">pBAD-RBS-Bovine Galectin-1-B0012 Terminator</span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">Soluble beta-galactoside binding protein (galectin) originally from Bos taurus. It is believed to expressed both intracellularly and extracellularly, where they serve to binding various types of beta-galactosides and initialize signal cascades. It is believed to be a dimer in its active form, as is true with most mammalian galectins.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">This biobrick is under the pBAD promoter (BBa_K206000) with attached RBS (BBa_B0034), and utilizes an additional RNA polymerase terminator (BBa_B0012).</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">UMaryland 2014 is interested in utilizing this part to investigate whether E. coli can recognize and bind to surface carbohydrates on a marine pathogen. Other uses of this part lie in the recognition of various carbohydrate ligands and potential activation of signal transduction pathways.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;"> </p><br />
<p style="text-align: center;"> </p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="text-align: left; line-height: 17.9400005340576px; white-space: pre-wrap;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; text-decoration: underline;">Part:BBa_K1489004 | </span></span><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; line-height: 17.9400005340576px; white-space: pre-wrap; text-decoration: underline;">pBAD-RBS-Lpp-OmpA-Bovine Galectin 1-B0012 Terminator</span></strong></span></p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; line-height: 17.9400005340576px; white-space: pre-wrap; text-decoration: underline;"><img src="https://static.igem.org/mediawiki/parts/0/0f/PSB1C3_-_BBa_K1489004.png" border="0" width="515" height="335" /></span></strong></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">His-tagged bovine galectin 1 attached to the c-terminus of OmpA, expressed under arabinose induction</span></span></p><br />
<p> </p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 20px; font-weight: bold; line-height: 19.5px; text-align: start;">pBAD-RBS-OmpA-Bovine Galectin-1-B0012 Terminator</span></span></p><br />
<p style="text-align: left;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 13px; line-height: 19.5px; text-align: start;">A fusion protein of OmpA (BBa_K1489002) fused to Bovine Galectin-1 (BBa_K1489001). Used to bind sugars at the outer cell membrane. Under pBAD regulation.</span></span></p><br />
<p style="text-align: left;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"> </span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"><span style="text-decoration: underline;"><strong>Part:BBa_K1489003 | Revised Lpp-OmpA</strong></span></span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"><span style="text-decoration: underline;"><strong><img src="https://static.igem.org/mediawiki/parts/9/90/PSB1C3_-_BBa_K1489003.png" border="0" width="515" height="335" /></strong></span></span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"> Biobrick 1, with a SacI site at the C-terminus replaced by a KasI site. This improvement was made in order to re-enable RE cloning; a SacI site exists in the chloramphenicol resistance gene.</span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 20px; font-weight: bold; line-height: 19.5px; text-align: start;">Lpp-OmpA-Linker with KasI site</span></span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">Outer membrane protein A (OmpA) is a common outer membrane protein found in bacteria that serves various purposes. For our purposes, the beta-barrel transmembrane domain can be utilized to anchor otherwise soluble proteins in the outer cell membrane. Lpp serves to direct the fusion protein, while a unstructured linker (GGGSGGGS) serves to separate OmpA from its cargo in order to avoid interactions between the two proteins.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">The expressed protein is almost structurally identical to BBa_K103006, with the terminal serine on the linker replaced by an alanine. This change was made in order to convert the previous SacI site (GAGCTC), which was unusable for RE cloning due to a second SacI site being present in the plasmid backbone, into a KasI site (GGCGCC). The KasI site codes for Gly-Ala, while the SacI site codes for Gly-Ser. This amino acid substitution is unlikely to impact the function of Lpp-OmpA-Linker in a major way.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;"> </p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;"> </p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">Part:BBa_K1489001 | Bovine Galectin-1</span></span></strong></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/parts/4/4c/PSB1C3_-_BBa_K1489001.png" border="0" width="515" height="335" style="border: 0;" /></span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">This is the Bovine Galectin which should interact with beta-galactosides and intialize a signal cascase</span></span></p><br />
<p style="text-align: center;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 20px; font-weight: bold; line-height: 19.5px; text-align: start;">Bovine Galectin-1</span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">Soluble beta-galactoside binding protein (galectin) originally from Bos taurus. It is believed to expressed both intracellularly and extracellularly, where they serve to binding various types of beta-galactosides and initialize signal cascades. It is believed to be a dimer in its active form, as is true with most mammalian galectins.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">UMaryland 2014 is interested in utilizing this part to investigate whether E. coli can recognize and bind to surface carbohydrates on a marine pathogen. Other uses of this part lie in the recognition of various carbohydrate ligands and potential activation of signal transduction pathways.</p><br />
<p style="margin: 0.4em 0px 0.5em; text-align: center;"><strong><span style="color: #282828; font-family: arial, sans-serif; font-size: small;"><span style="line-height: 19.5px;">Part:BBa_K1489005: </span></span><span style="text-align: left; line-height: 19.5px;"><span style="color: #282828; font-family: arial, sans-serif; font-size: small;">Crasseoterra virginica Galectin 1 (CvGal1)</span></span></strong></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;"> </p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;"><img src="https://static.igem.org/mediawiki/parts/0/01/UM_PSB1C3_-_BBa_K1489005.png" border="0" width="515" height="335" style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; line-height: 17.9400005340576px; text-align: center; white-space: pre-wrap; cursor: se-resize !important; display: block; border: 0px; margin-left: auto; margin-right: auto;" /></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px; text-align: center;"><span style="font-size: 20px; font-weight: bold; text-align: start;">Crasseoterra virginica Galectin 1 (CvGal1)</span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;"><span style="font-size: 13px;">His-tagged soluble beta-galactoside binding protein (galectin) originally from Crasseoterra virginica. CvGal1 is believed to expressed both intracellularly and extracellularly, where they serve to binding various types of n-acetylgalactosamines and initialize signal cascades. It is also believed to be a tetramer in its active form, as is true with most invertebrate galectins.</span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;"><span style="font-size: 13px;"> </span></p><br />
<p style="margin: 0.4em 0px 0.5em; text-align: center;"><span style="text-decoration: underline;"><strong><span style="color: #282828; font-family: arial, sans-serif; font-size: small; text-decoration: underline;"><span style="line-height: 19.5px;">Part:BBa_K1489006: </span></span><span style="color: #282828; font-family: arial, sans-serif; font-size: small;"><span style="line-height: 19.5px;">OmpA-CvGal1</span></span></strong></span></p><br />
<p style="margin: 0.4em 0px 0.5em; text-align: center;"><span style="text-decoration: underline;"><strong><span style="color: #282828; font-family: arial, sans-serif; font-size: small;"><span style="line-height: 19.5px;"> </span></span></strong></span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;"><img src="https://static.igem.org/mediawiki/parts/2/28/UM_PSB1C3_-_BBa_K1489006.png" border="0" width="515" height="335" style="font-size: small; color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; line-height: 17.9400005340576px; text-align: center; white-space: pre-wrap; display: block; cursor: se-resize !important; border: 0px; margin-left: auto; margin-right: auto;" /></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px; text-align: center;"><span style="font-size: 20px; font-weight: bold; text-align: start;">OmpA-CvGal1</span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px; text-align: center;"><span style="font-size: 20px; font-weight: bold; text-align: start;"> </span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px; text-align: left;"><span style="font-size: 13px;">CvGal1 fused to OmpA in order to anchor it in the membrane.</span></p> <br />
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<option value="https://2014.igem.org/Team:UMaryland/meetourteam">&nbsp;--Meet Our Team Members</option><br />
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Team:UMaryland/humanprac/publicawareness
2014-10-17T19:46:16Z
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<h1 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/humanprac/publicawareness" class="contentpagetitle"> <br />
Public Awareness <br />
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<p class="MsoNormal" style="text-align: center;"><strong><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"><span style="text-decoration: underline;">Public Awareness</span></span></strong></p><br />
<p> </p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">Realizing the importance of education and understanding, we strived to improve general understanding of <em>P. marinus</em> and the Dermo infection. We began at the University of Maryland with a survey to get students from all backgrounds to start thinking about the problems we were addressing. Next, we publicized our project and goals by connecting with deans of various colleges to spark interest in these fields. As awareness increased on campus, we turned our attention to the surrounding community. Networking with schools around the area, programs were established where iGEM members would go to the school and give a presentation or seminar tailored to the interests of the audience. For example, we gave a seminar on biotechnology and synthetic biology at a local high school in order to start education early with students who will be at the forefront of this type of research in the near future. The next step will be to get in touch with state organizations such as the Chesapeake Bay foundation in order to devise outreach initiatives and progressively raise awareness at higher levels of the community.</span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">Additionally we conducted a survey of over 250 students in undergraduate microbiology classes at the University of Maryland to further understand the public perception of this technology through the eyes of our next generation.The survey and responses are found below.</span></p><br />
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<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"> </span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">All responses to the survey will be kept anonymous, and no personal information about the participants will be released. The results of the survey may be published.</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;"> </span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">I have read and consent to the terms of confidentiality listed above ___________</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">1) What is a biosensor?</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">a. an organ that measures biological phenomenon</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">b. an organism that has the ability to sense other organisms</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">c. a device that measures biological parameters such as temperature or pressure</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><strong><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">d. an instrument that has a biological component and measures physical or chemical parameters</span></strong></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><strong><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;"> </span></strong></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">2) How do you feel about genetic engineering?</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">a. Strongly Support</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">b. Support</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">c. Neutral</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">d. Oppose</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">e. Strongly Oppose</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;"> </span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">3) How do you feel about the introduction of genetically modified </span><em style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">E. Coli</em><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;"> into the Chesapeake Bay?</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">a. Strongly Support</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">b. Support</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">c. Neutral</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">d. Oppose</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">e. Strongly Oppose</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;"> </span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">4) If it improved Chesapeake bay health and aquaculture?</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">a. Strongly Support</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">b. Support</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">c. Neutral</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">d. Oppose</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">e. Strongly Oppose</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;"> </span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">5) What danger do you think lies in using biotechnology and bioengineering to address environmental issues?</span></p><br />
<p class="MsoNormal"> </p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">(open ended)</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; text-align: center; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="text-decoration: underline;"><strong><span style="font-size: 10pt; font-family: Arial, sans-serif; color: #222222; text-decoration: underline;">ANSWER</span></strong></span><strong><span style="font-size: 10pt; font-family: Arial, sans-serif; color: #222222; text-decoration: underline;">S</span></strong></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;"><img src="https://static.igem.org/mediawiki/2014/1/15/UMSurveyAnswer1.jpg" border="0" width="365" height="268" style="float: left;" /></span><strong style="text-align: center;"><span style="font-size: 10pt; font-family: Arial, sans-serif; color: #222222; text-decoration: underline;"><img src="https://static.igem.org/mediawiki/2014/e/e9/UMSurveyAnswer2.jpg" border="0" width="410" height="268" style="cursor: se-resize !important; border: 0;" /></span></strong></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"> </p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><img src="https://static.igem.org/mediawiki/2014/0/07/UMSurveyAnswer3.jpg" border="0" width="365" height="268" style="border: 0;" /><img src="https://static.igem.org/mediawiki/2014/d/d2/UMSurveyAnswer4.jpg" border="0" width="365" height="268" /></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">By administering this survey to an undergraduate general microbiology class at the University of Maryland, we hoped to gain insight about where students stood on genetic engineering and its applications. We chose this class because it is a general education class at the University of Maryland which attracts students of various backgrounds and majors, not just science majors.</span></p><br />
<p class="MsoNormal"><span style="font-family: 'Times New Roman', serif; font-size: 12pt; line-height: 115%;"> With the first question, we tried to figure out just what proportion of the students were science majors or non-science majors, so that we could better interpret the responses to the later questions. We chose to ask what a biosensor was to determine if the student had significant background in biology or perhaps exposure to biotechnology. To our surprise, most students answered this question correctly which suggests that many students probably had some experience with or have heard about aspects of bioengineering. This conclusion reveals the extent of Science, Technology, Engineering, and Mathematics (STEM) education of University of Maryland students and the fact that most are fairly well informed about this type of technology.</span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">The next three questions corroborate the results from first question as shown by a relatively high number of students who are open to the idea of genetic engineering of <em>E. coli</em>. Even after being asked bluntly about the introduction of genetically-altered <em>E. coli</em> into the bay without any other information, more students, surprisingly, remained neutral rather than opposed. Finally, as expected, after qualifying the previous question by providing the motivation for the use of <em>E. coli</em>, a large shift was observed towards being supportive of the proposition. </span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"> Lastly, the free response questions (responses not shown) provided a deeper insight into the understanding of the population being surveyed. Most of the students developed valid points regarding possible areas of concern with this type of project. These ranged from ethical concerns to long-term, unexpected problems and the idea that native species may be adversely affected by our biosensor. It seemed to us that most of the concerns stemmed from a lack of understanding of the technology being used. Education and outreach efforts would likely eliminate some of these misunderstandings; however, some of the concerns raised are valid and are points we considered ourselves.</span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">The next step in this project would be to broaden the population being surveyed both in terms of size and diversity. In this way, we can determine if the results we obtained here is actually representative of a university setting. In terms of the participants being surveyed, we believe that young students are the most important because this is the generation that will be deciding on novel technologies that address pressing social and environmental concerns such as the health of the Chesapeake Bay. As a result, it is important to ensure that this demographic, especially, has access to correct information so that informed decisions can be made. However, we acknowledge that it is also important to gauge the views of current policy makers and people outside the university as well, so future surveys may be targeted at older demographics and different regions.</span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">With regards to the effect on public outreach efforts, it is clear that although most of the responses appeared to be informed there is room for improvement. Accordingly, we plan to continue our on campus and community efforts to advertise the issue and our research in order to start a thoughtful dialogue between those affected by concerns regarding the Chesapeake Bay and those who are trying to address these concerns.</span></p><br />
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Team:UMaryland/humanprac/community
2014-10-17T19:45:54Z
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<p class="MsoNormal" style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">Interaction with Community</span></strong></span></p><br />
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<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">We primarily focused our efforts on three organizations in the Chesapeake Bay community: Marinetics Choptank Oyster Farm, Department of Natural Resources, and Chesapeake Bay Foundation. Each provided insight into a different dimension of the Dermo infection problem. At Marinetics, we received an economy-centric view, where we learned a great deal about how <em>P. marinus</em> actually affects commercial oyster growth. We were surprised to find that, at least in that region of the bay, the owner of the farm experienced little to no effect from the pathogen. However, he did concede that detection systems for Dermo are not adequate and expressed that a real-time biosensor would be quite useful.</span></p><br />
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<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"><img src="https://static.igem.org/mediawiki/parts/c/c8/Oysterfarm2.jpg" border="0" width="319" height="180" style="display: block; margin-left: auto; margin-right: auto;" /></span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">Next, we explored <em>P. marinus</em> through an environmental perspective as we spoke to members of the Chesapeake Bay Foundation. Here, we heard what was more expected of the problem including its adverse effects on oyster populations in the Chesapeake Bay. This interaction revealed the larger impacts our research would have on bay health as well as informed us about current approaches to remedy the problem. Still, we were unsure what to make of the two seemingly opposite views presented by the oyster farm and the Chesapeake Bay Foundation.</span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"> </span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"><img src="https://static.igem.org/mediawiki/parts/1/15/UMDNR2.jpg" border="0" width="180" height="241" style="display: block; margin-left: auto; margin-right: auto;" /></span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">Lastly, to clarify techniques and detection methods, we arranged a meeting with staff and lab members from the Department of Natural Resources. Mr. Chris Dungan, who is in charge of <em>P. marinus</em> investigation among other pathogens, gave us a comprehensive overview of how Dermo infections are detected and handled. We also discussed different aspects of policy-making and the obstacles to introducing new technology to combat these problems. For example, Mr. Dungan reconciled the apparently opposing accounts we received from the Chesapeake Bay Foundation and the oyster farm by telling us that Dermo infection is more prevalent in certain areas of the bay and will adversely affect populations in the bay but maybe not in aquaculture because oysters would be harvested and cooked before they exhibited the pathophysiology of the disease. Moreover, he acknowledged that oyster farmers tend to pay less attention to <em>P. marinus</em> in general because it is not a human pathogen. </span></p><br />
<p> </p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">Ultimately, the fact that people tend to be apprehensive about new technologies and research that could be used to solve environmental problems motivated us to help address this issue. Since we understood just how powerful public opinion can be in promoting or labeling potential solutions prematurely as dangerous, we developed two main initiatives to promote education in these technologies and to gauge the public opinion quantitatively in our local community to understand why there is such reluctance towards new technology and the reasons behind this.</span></p> <br />
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<option value="https://2014.igem.org/Team:UMaryland">&nbsp;Homepage</option><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/humanprac/publicschool
Team:UMaryland/humanprac/publicschool
2014-10-17T19:45:03Z
<p>Nbb826: </p>
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School Outreach <br />
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<br />
<p> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-align: center;" dir="ltr"><span style="text-decoration: underline;"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: underline; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">Churchill High School</span></strong></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="text-decoration: underline;"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: underline; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;"> </span></strong></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="text-decoration: underline;"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: underline; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;"> </span></strong></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-align: center;" dir="ltr"><span style="text-decoration: underline;"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: underline; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;"><img src="https://static.igem.org/mediawiki/parts/b/bb/Pavelschoolvisit.jpg" border="0" width="494" height="371" style="border: 0;" /><br /></span></strong></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">In June of 2014, a member of our group conducted a short presentation about genetic engineering for an AP Biology class at Winston Churchill High School in Potomac, Maryland. The presentation contained information on the process of genetic engineering in general including transcription and translation, the basics of bacterial genetics, plasmid design; and on our team’s project including the importance of oysters to the Chesapeake Bay ecosystem and economy, the mechanism by which </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> infects oysters, and the protein structure being designed for our project. Approximately half of the class period, both during the presentation and after it, was left for students to ask questions. Some of the students were clearly interested in learning more about the current and future applications of genetic engineering, such as in agriculture, and about the possible risks involved and how they can be reduced. Some of the students asking these questions later said after the presentation that they were interested in studying genetics in college. Overall, the presentation was understandable and engaging to most of the class and several students were clearly interested in learning more about IGEM and our team’s current project.</span></p> <br />
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<div class="yjsg7_before"><div id="yjsg7" class="yjsg_grid yjsgsitew"><div id="user23" class="yjsgxhtml only_mod"><div class="yjsquare modid117"><div class="h2_holder"><h2 class="module_title"><span class="title_split titlesplit0">About</span> <span class="title_split titlesplit1">Umaryland</span></h2></div><div class="yjsquare_in"><p>UMaryland 2014 is the inaugural iGEM team of the University of Maryland, College Park. We are a combined effort of several departments and numerous faculty mentors. Although it is only our first year, we believe our hard work and dedication has paid off. We can't wait for this year's competition! GO TERPS!</p></div></div></div></div></div> <div class="footer_holders footer"><!-- footer --><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/humanprac/overview
Team:UMaryland/humanprac/overview
2014-10-17T19:44:04Z
<p>Nbb826: </p>
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<li class="item101 level0 first"><span class="mymarg"><a class="yjanchor first" href="https://2014.igem.org/Team:UMaryland"><span class="yjm_has_none"><span class="yjm_title">Homepage</span></span></a></span></li><li class="item104 level0"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/photos"><span class="yjm_has_none"><span class="yjm_title">Photos</span></span></a></span></li><li class="haschild item105 level0"><span class="child"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/meetourteam"><span class="yjm_has_none"><span class="yjm_title">Team</span></span></a></span><ul class="subul_main level1"><li class="bl"></li><li class="tl"></li><li class="tr"></li><li class="item293 level1 first"><span class="mymarg"><a class="yjanchor first" href="https://2014.igem.org/Team:UMaryland/meetourteam"><span class="yjm_has_none"><span class="yjm_title">Meet Our Team Members</span></span></a></span></li><li class="item116 level1"><span 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href="https://2014.igem.org/Team:UMaryland/project/futureapplications"><span class="yjm_has_none"><span class="yjm_title">future applications</span></span></a></span></li><li class="item301 level1 lilast"><span class="mymarg"><a class="yjanchor last" href="https://igem.org/Safety/Safety_Form?team_id=1489"><span class="yjm_has_none"><span class="yjm_title">Safety</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li><li class="haschild item234 level0"><span class="child"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/notebook"><span class="yjm_has_none"><span class="yjm_title">Notebook</span></span></a></span><ul class="subul_main level1"><li class="bl"></li><li class="tl"></li><li class="tr"></li><li class="item303 level1 first lilast"><span class="mymarg"><a class="yjanchor firstlast" href="https://2014.igem.org/Team:UMaryland/notebook"><span class="yjm_has_none"><span class="yjm_title">Wetlab Notebook</span></span></a></span></li><li 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Overviewmissionstatement <br />
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<p class="MsoNormal" style="text-align: center;"><strong><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"><span style="text-decoration: underline;">Overview Mission Statement</span></span></strong></p><br />
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<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">From the beginning, our team wanted to pursue a project that would represent the spirit of Maryland. What better than to turn to the Chesapeake Bay, a natural resource that is priceless to Marylandians. The Chesapeake Bay, in addition to being a beautiful attraction, supports aquaculture and ecosystems that are essential to the economy and the environment. So, after choosing to address an oyster pathogen, one of many issues that threaten Chesapeake Bay health, we sought to understand the implications of our project and how it could be applied.</span></p><br />
<p> </p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">We organized various meetings with different Chesapeake Bay organizations to gauge the usefulness of our research and to further optimize our design. In doing so, we were made aware of the fact that public opinion played a large part in governing what research would be admissible and what the final product of the research could look like. Then, we took it to the next step to start our own initiatives to help educate and spread awareness about synthetic biology, and iGEM specifically in the context of the Chesapeake Bay and the environment. This outreach was accomplished through surveying a population of students and giving a class for middle school students.</span></p> <br />
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Team:UMaryland/project/futureapplications
2014-10-17T19:43:11Z
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<a href="https://2014.igem.org/Team:UMaryland/project/futureapplications" class="contentpagetitle"> <br />
Future Applications <br />
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<p>Future Applications of Research</p><br />
<p>We believe that the initial stages of the Biosensor have been complete and will lead to many new and exciting developments in the future. The general idea is that once the surface protein has been expressed, it will be able to interact with Dermo and produce a signal.</p><br />
<p>As for this year we our efforts working with a Bovine-Galectin have come the furthest and are ready to move into these next steps. Additionally in future years we hope to incorporate CvGal1 into the constructs, an effort we were not able to accomplish due to lack of time and troubleshooting. </p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong>Signaling Approaches:</strong></span></p><br />
<p> </p><br />
<p>1). Genetic engineering approach -Downstream Signaling Cascade</p><br />
<p><img src="https://static.igem.org/mediawiki/parts/9/99/UMSIgnal_Cascade.jpg" border="0" width="671" height="503" style="line-height: 12.0000009536743px; display: block; margin-left: auto; margin-right: auto; cursor: se-resize !important;" /></p><br />
<p>One of the strategies we hope to pursue is developing or incorporating a downstream signaling cascade into the cell strain. In this effort, once the E. coli came in contact with the Dermo it would then set off a transcription factor, ultimately producing a signal. Multiple approaches have been researched, including a Cpx membrane stress cascade, a split GFP cascade, and others.</p><br />
<p>The Cpx two component system reacts naturally to membrane stress. When the much larger Dermo is bound to the surface receptor expressed on E. coli, the bacterial cell membrane would become deformed. In turn, this would transmit a signal to the nucleus via the NlpE system that could be coupled to GFP production. This assay would require florescence microscopy to detect Dermo in solution.</p><br />
<p>Another option is for a split GFP construct. The idea is two different constructs containing the ligand are present in the cell, each with a different fraction of the GFP gene. When Dermo comes in close contact with the cell, multiple ligands on the cell would bind to the receptors on Dermo, causing the membrane to pinch or come together. This would trigger a spontaneous chance that two complementary pieces of GFP in close contact would bind, producing a fluorescent signal. This would also need to be observed using microscopy.</p><br />
<p>One of the ultimate goals from using the downstream signaling approach is to develop a “Kamikaze Bacteria”. By this we hope that the bacteria would respond to the presence of Dermo by eliminating it, possibly through killing itself. Similar work has previously been done with bees and colony collapse.</p><br />
<p>2). Microfluidics Approach- Size Based Cell sorter</p><br />
<p style="text-align: center;"><img src="https://static.igem.org/mediawiki/parts/0/02/UMMicrofluidics.png" border="0" width="572" height="416" /></p><br />
<p>Another avenue we plan to pursue is a microfluidics based detection approach. The overall concept would rely on the fact that E.coli cells are much smaller than Dermo cells. By creating a microfluidic device that sorts based on size, E.coli would travel into one output, whereas the Dermo and Dermo+E.coli complexes would travel into another output. Furthermore if the E.coli cells are fluorescently stained, analysis of the larger size fraction would indicate whether any E.coli cells are bound, confirming the presence of Dermo. While further differentiations based on size between bound and unbound Dermo may not be possible due to growth differences, this method will still suffice. </p><br />
<p>A microfluidic device would also contain the E.coli preventing them from entering the surrounding environment. Containment would be an obvious concern if a device like this were to ever hit the market. In this solution theoretically one could have pre-labeled E.coli in the machine stored and ready. The end user would simply intake a water sample right from the source, allow it to incubate, and process (sort) it. Collecting the larger fraction and viewing under a microscope would give a quick and easy real time method for detecting Dermo.</p><br />
<p>We believe that with the groundwork we have laid this year these future endeavors will be made possible as UMaryland continues to grow and gain expertise.</p><br />
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<div class="yjsg7_before"><div id="yjsg7" class="yjsg_grid yjsgsitew"><div id="user23" class="yjsgxhtml only_mod"><div class="yjsquare modid117"><div class="h2_holder"><h2 class="module_title"><span class="title_split titlesplit0">About</span> <span class="title_split titlesplit1">Umaryland</span></h2></div><div class="yjsquare_in"><p>UMaryland 2014 is the inaugural iGEM team of the University of Maryland, College Park. We are a combined effort of several departments and numerous faculty mentors. Although it is only our first year, we believe our hard work and dedication has paid off. We can't wait for this year's competition! GO TERPS!</p></div></div></div></div></div> <div class="footer_holders footer"><!-- footer --><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/project/results
Team:UMaryland/project/results
2014-10-17T19:42:21Z
<p>Nbb826: </p>
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<a href="https://2014.igem.org/Team:UMaryland/project/results" class="contentpagetitle"> <br />
Results <br />
</a><br />
</h1><br />
<br />
<br />
<br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/2014/e/e0/UMFig1-1.jpg" border="0" width="213px;" height="248px;" style="border: NaNpx solid black; transform: rotate(0.00rad); -webkit-transform: rotate(0.00rad);" /></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/2014/2/28/UMFig1-2.png" border="0" width="354px;" height="148px;" style="border: NaNpx solid black; transform: rotate(0.00rad); -webkit-transform: rotate(0.00rad);" /></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> A B</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig. 1 – A: Double RE digest (EcoRI and PstI) of pBAD-Bovine Galectin 1 and OmpA-Bovine Gibson assemblies. For pBAD-BtGal1, all lanes for after ladder have bands of the correct weight, while no lanes for OmpA-Bovine are of the correct weight. B: Expected band sizes after double digest for pBAD-Bovine Galectin 1. The EcoRI cut site in the Bovine Galectin 1 gene was subsequently removed via site directed mutagenesis.</span></p><br />
<p><strong id="docs-internal-guid-809a97bc-f600-9f66-fada-73accace1d98" style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Mammalian galectins typically have a simpler quaternary structure than invertebrate galectins; they are usually dimers while invertebrate galectins are usually tetramers. Bovine galectin 1, from Bos taurus, is a simpler galectin than CvGal1 and can thus be utilized as a model galectin to anchor in the E. coli outer membrane. In addition, while the crystal structure of CvGal1 and the exact binding ligand on P. marinus that CvGal1 binds to are still under investigation, the crystal structure and nature of bovine galectin 1 have been well studied. We have cloned Bovine galectin 1 into the pSB1C3 backbone via Gibson assembly. The bovine galectin 1 gene was ordered as a gBlock from IDT and PCR amplified for Gibson assembly. All Gibson products were verified via sequencing. </span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/2014/3/3f/UMFig2-1.jpg" border="0" width="233" height="331" style="border: 0;" />A<img src="https://static.igem.org/mediawiki/2014/e/ef/UMFig2-2.png" border="0" width="354px;" height="151px;" style="border: 0; transform: rotate(0.00rad); -webkit-transform: rotate(0.00rad);" /></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">B</span></p><br />
<p><strong style="font-weight: normal;"> (3rd Lane contains OmpA-Bovine product)</strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig. 2 - </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">OmpA-Bovine (cut with BamHI and HindIII, before SDM)</span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig. 2 – Double digest of OmpA-Bovine Galectin 1 (BamHI and HindIII) Lane 3 contains bands of the correct weight after digest B: Expected band sizes after double digest for pBAD-Bovine Galectin 1. Site directed mutagenesis was subsequently performed in order to remove an EcoRI site in the Bovine galectin 1 gene. Figure </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">2A</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> shows the Bovine galectin-1 assembled into pSB1C3-pBAD-RBS as a fusion protein with OmpA. When expressed in </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">E. coli</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">, the galectin should continue off the C-terminus of the OmpA-linker and thus be transported to the extracellular matrix since it is attached to the OmpA transport protein. After plasmid assembly transfer of plasmid into a DH5α </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">E. coli</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> cells strain, the plasmid was extracted and digested with EcoRI and PstI. The pattern of three bands appears again due to an additional EcoRI restriction site. </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 12.0000009536743px;"><span><img src="https://static.igem.org/mediawiki/2014/4/41/UMFig4-1.jpg" border="0" width="735" height="272" /></span></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 12.0000009536743px;">Sequencing Results:</p><br />
<p style="line-height: 12.0000009536743px;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$0:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$0:0">OmpA-Bovine SDM Chromatogram:</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$1:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$2:0">Sequence file:</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$3:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$4:0">NNNNNNNNNNNTNNNNANNNNNAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGGATG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$5:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$6:0">ATTTCTGGAATTCGCGGCCGCTTCTAGAGACATTGATTATTTGCACGGCGTCACACTTTGCTATGCCATAGCAAGATAGT</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$7:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$8:0">CCATAAGATTAGCGGATCCTACCTGACGCTTTTTATCGCAACTCTCTACTGTTTCTCCATACCGTTTTTTTGGGCTAGCT</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$9:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$10:0">ACTAGAGAAAGAGGAGAAATACTAGCATATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGC</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$11:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$12:0">TGGCAGGTTGCTCCAGCAACGCTAAAATCGATCAGGGAATTAACCCGTATGTTGGCTTTGAAATGGGTTACGACTGGTTA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$13:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$14:0">GGTCGTATGCCGTACAAAGGCAGCGTTGAAAACGGTGCATACAAAGCTCAGGGCGTTCAACTGACCGCTAAACTGGGTTA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$15:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$16:0">CCCAATCACTGACGACCTGGACATCTACACTCGTCTGGGTGGCATGGTATGGCGTGCAGACACTAAATCCAACGTTTATG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$17:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$18:0">GTAAAAACCACGACACCGGCGTTTCTCCGGTCTTCGCTGGCGGTGTTGAGTACGCGATCACTCCTGAAATCGCTACCCGT</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$19:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$20:0">CTGGAATACCAGTGGACCAACAACATCGGTGACGCACACACCATCGGCACTCGTCCGGACAACGGCGGANGTTCTGGAGG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$21:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$22:0">ANGGAGCATGGCGTCGGGGCTTGTTGCGTCCAATCTGAATCTGAAACCAGGTGAAAGCTTACGCGTCCGCGGCGAAGTAG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$23:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$24:0">CCGCAGATGCGAAATCTTTTCTCTTGAACCTGGGCAAGGACGANAATAATCTGGCTTTGCATTTCAACCCGCGTTTCNAC</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$25:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$26:0">GCTCATGGNGACNTGAACACAATCNTGTGTAANTCCAAGGNTGCTGGTGCTTGGGGTGCANAANNANCGNGNNNTCCGCG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$27:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$28:0">TTCCCGTTTCANNNNNGCTCANNNNGGNAGTCACCNTNCNGCNTTCANCCANNNCGANNTNCNNNNTTAAANNNGNNNNN</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$29:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$30:0">NGGNNNCNAAGTTNNANTTNNCNAANCNNNTNNAACNNNNNGNNNNNNNNNNNNNNNNNGNNNN</span></span></p><br />
<p style="line-height: 12.0000009536743px;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$0:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$30:0">A</span></span></p><br />
<p style="line-height: 12.0000009536743px;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$0:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$30:0"><img src="https://static.igem.org/mediawiki/2014/b/bc/UMFig5-1.jpg" border="0" width="733" height="272" /></span></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"> </p><br />
<p style="line-height: 12.0000009536743px;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$0:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$30:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$0:0">pBAD-Bovine SDM chromatogram:</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$1:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$2:0">NNNNNNNNNNNNTNNNCTATNNNNTAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$3:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$4:0">ATGATTTCTGGAATTCGCGGCCGCTTCTAGAGACATTGATTATTTGCACGGCGTCACACTTTGCTATGCCATAGCAAGAT</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$5:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$6:0">AGTCCATAAGATTAGCGGATCCTACCTGACGCTTTTTATCGCAACTCTCTACTGTTTCTCCATACCGTTTTTTTGGGCTA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$7:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$8:0">GCTACTAGAGAAAGAGGAGAAATACTAGATGGCGTCGGGGCTTGTTGCGTCCAATCTGAATCTGAAACCAGGTGAAAGCT</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$9:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$10:0">TACGCGTCCGCGGCGAAGTAGCCGCAGATGCGAAATCTTTTCTCTTGAACCTGGGCAAGGACGATAATAATCTGGCTTTG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$11:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$12:0">CATTTCAACCCGCGTTTCAACGCTCATGGCGACGTGAACACAATCGTGTGTAATTCCAAGGATGCTGGTGCTTGGGGTGC</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$13:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$14:0">AGAACAGCGTGAGTCCGCGTTCCCGTTTCAACCGGGCTCAGTTGTGGAAGTCACCATCAGCTTCAACCAGACCGATCTCA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$15:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$16:0">CCATTAAACTGCCGGATGGCTACGAGTTTAAGTTTCCAAACCGTTTAAACCTCGAGGCCATCAACTATCTGTCCGCGGGG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$17:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$18:0">GGCGACTTCAAGATTAAAAGCGTGCACCATCATCACCACCATTGATCACACTGGCTCACCTTCGGGTGGGCCTTTCTGCG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$19:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$20:0">TTTATATACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTTCTTTAAAACCGA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$21:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$22:0">AAAGATTACTTCGCGTTATGCNGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$23:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$24:0">GCTCACTCAAAGGCGGTAATACGGTTATCNNCAGAATCNGGGGATNACGCAGGAAANAACATGTGAGCAAANNNCAGCAA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$25:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$26:0">AANGCAGGAANCGTAAAANNNN</span></span></span></p><br />
<p style="line-height: 12.0000009536743px;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$0:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$30:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$26:0">B</span></span></span></p><br />
<p> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig. 3 - <strong>S</strong></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">equencing results and Chromatogram of SDM’d plasmids (</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">BBa_K1489000 and BBa_K1489004)</span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">To address this issue, site-directed mutagenesis was performed on these plasmids to change the EcoRI sequence within the bovine galectin-1 gene (5’-GAATTC-3’) to 5’-GAGTTT-3’, which will code for the same Glu-Phe sequence. Figure </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">3A</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> and </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">3B</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> demonstrate the two successfully mutagenized base pairs in the bovine galectin-1 gene, while preserving the rest of the plasmid.</span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p> <img src="https://static.igem.org/mediawiki/parts/a/a5/UMFinalfigureigem.jpg" border="0" width="687" height="269" style="line-height: 12.0000009536743px; cursor: se-resize !important;" /></p><br />
<p> Sequencing results:<br />
NNNNNNNNNNNNNNNNTATANAAATAGGCGTATCNCGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGG<br />
ATGATTTCTGGAATTCGCGGCCGCTTCTAGAGGGCATATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTT<br />
CTACTCTGCTGGCAGGTTGCTCCAGCAACGCTAAAATCGATCAGGGAATTAACCCGTATGTTGGCTTTGAAATGGGTTAC<br />
GACTGGTTAGGTCGTATGCCGTACAAAGGCAGCGTTGAAAACGGTGCATACAAAGCTCAGGGCGTTCAACTGACCGCTAA<br />
ACTGGGTTACCCAATCACTGACGACCTGGACATCTACACTCGTCTGGGTGGCATGGTATGGCGTGCAGACACTAAATCCA<br />
ACGTTTATGGTAAAAACCACGACACCGGCGTTTCTCCGGTCTTCGCTGGCGGTGTTGAGTACGCGATCACTCCTGAAATC<br />
GCTACCCGTCTGGAATACCAGTGGACCAACAACATCGGTGACGCACACACCATCGGCACTCGTCCGGACAACGGCGGAGG<br />
TTCTGGAGGAGGGAGCTCTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTT<br />
CTTTAAAACCGAAAAGATTACTTCGCGTTATGCAGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGG<br />
CGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCANGNNNAACATGTGAGCA<br />
AAAGGCCAGCNNAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCACAGGCTCCGCCCCCCTGACGAGCAT<br />
CACAAAAATCGACGCTCAAGTCAGANGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGNAGCTC<br />
CCTCGTGCGCTCTCCTGTTCCNACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTT<br />
CTCNTAGCTCACGCTGTAGGTATCNNCANNN</p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig 4. - <strong>S</strong></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">equencing Results and Chromatogram of OmpA moved into pSB1C3</span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Since UMaryland worked extensively with the OmpA-linker (BBa_K103006), we have also looked into improvements on that BioBrick.</span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; background-color: #f7f7f7;">OmpA-KasI chromatogram, shrunk down</span></span></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 12.0000009536743px;"><img src="https://static.igem.org/mediawiki/2014/0/0b/UMFig3-1.jpg" border="0" width="687" height="269" /></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 12.0000009536743px;">Sequencing Results:</p><br />
<p style="line-height: 12.0000009536743px;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$4:0">Sequence:</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$5:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$6:0">NNNNNNNNNNNNNNNTATAAAAATAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGGA</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$7:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$8:0">TGATTTCTGGAATTCGCGGCCGCTTCTAGAGGGCATATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTC</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$9:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$10:0">TACTCTGCTGGCAGGTTGCTCCAGCAACGCTAAAATCGATCAGGGAATTAACCCGTATGTTGGCTTTGAAATGGGTTACG</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$11:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$12:0">ACTGGTTAGGTCGTATGCCGTACAAAGGCAGCGTTGAAAACGGTGCATACAAAGCTCAGGGCGTTCAACTGACCGCTAAA</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$13:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$14:0">CTGGGTTACCCAATCACTGACGACCTGGACATCTACACTCGTCTGGGTGGCATGGTATGGCGTGCAGACACTAAATCCAA</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$15:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$16:0">CGTTTATGGTAAAAACCACGACACCGGCGTTTCTCCGGTCTTCGCTGGCGGTGTTGAGTACGCGATCACTCCTGAAATCG</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$17:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$18:0">CTACCCGTCTGGAATACCAGTGGACCAACAACATCGGTGACGCACACACCATCGGCACTCGTCCGGACAACGGCGGAGGT</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$19:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$20:0">TCTGGAGGAGGGGCGCCTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTTC</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$21:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$22:0">TTTAAAACCGAAAAGATTACTTCGCGTTATGCAGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGC</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$23:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$24:0">GAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCA</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$25:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$26:0">AAAGGNCAGCAAAAGGNCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCACAGGCTCCGCCCCCCTGACGAGCA</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$27:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$28:0">TCACAAAAATCGACGCTCAAGTCANNNNGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTC</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$29:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$30:0">CCTCGTGCGCTCTNCTGTTCCNACCCTGCCGCTTACCNGATACCTGTCNCNTTCTNCCTTCGGGANCGNGNGCTTTCTCA</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$31:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$32:0">TAGCTCACGCTNTAGGTANNNCANTNCGGNNNTNGNCGTTNNNCNCNANNCTNGNNTN</span></p><br />
<p> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig. 5 - <strong>S</strong></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">equencing Results and Chromatogram of SDM’d OmpA in pSB1C3</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><br /></span><span style="line-height: 12.777777671814px;">The previous version of Lpp-OmpA-Linker (BBa_K103006), designed for the pSB1A2 vector backbone, contained the non-traditional restriction sites NdeI (CATATG) and SacI (GAGCTC) in order to attach proteins at the N-terminus and C-terminus of OmpA, respectively. While the NdeI site is still useful for RE cloning at the N-terminus of OmpA, the implementation of the new plasmid backbone pSB1C3 has made the SacI site difficult to use. This is due to the presence of a second SacI site in the chloramphenicol acetyltransferase gene in pSB1C3. We have thus decided to mutagenize the previous SacI site on Lpp-OmpA-Linker into a KasI site in order to remove this issue. The KasI site (GGCGCC) encodes the amino acids glycine and alanine, converting the sequence of the linker from Gly-Gly-Gly-Ser-Gly-Gly-Gly-Ser to Gly-Gly-Gly-Ser-Gly-Gly-Gly-Ala. We believe this to be a sufficiently minor alteration to the unfolded linker sequence as to not affect protein function.</span></p> <br />
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<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; background-color: #f7f7f7;"></span></span></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 12.0000009536743px;"><img src="https://static.igem.org/mediawiki/parts/0/0c/UMBovinegalectinexpression.jpg" border="0" width="687" height="269" /></p><br />
<br />
<p> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig. 6 - <strong>S</strong></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Western Blot of expression of SDM’d OmpA-BovineGalectin in pSB1C3</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><br /></span><span style="line-height: 12.777777671814px;">"Fig. 6 - Expression test for pBAD-Bovine galectin 1 and OmpA-Bovine galectin 1. Western Blot with anti-His antibody was done on soluble and insoluble fractions of non-induced cells and induced cells (0.2% arabinose for 21 hours). Bands can be observed in induced cells for pBAD-Bovine galectin 1 and both induced and non-induced cells for OmpA-Bovine galectin 1"</span></p><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/project/biobricks
Team:UMaryland/project/biobricks
2014-10-17T19:41:56Z
<p>Nbb826: </p>
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<h1 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/project/biobricks " class="contentpagetitle"> <br />
Biobricks <br />
</a><br />
</h1><br />
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<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; text-decoration: underline;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">Part:BBa_K1489002 | </span></span><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap; text-decoration: underline;">Lpp-OmpA in new backbone</span></strong></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/parts/f/fd/PSB1C3_-_BBa_K1489002.png" border="0" width="480" height="335" style="border: 0;" /></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;">We moved an existing biobrick, BBa_K103006, from an old plasmid backbone, pSB1A2, into the current backbone, pSB1C3, via RE cloning.</span></p><br />
<p style="text-align: center;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 20px; font-weight: bold; line-height: 19.5px; text-align: start;">Lpp-OmpA-Linker (pSB1C3)</span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">Outer membrane protein A (OmpA) is a common outer membrane protein found in bacteria that serves various purposes. For our purposes, the beta-barrel transmembrane domain can be utilized to anchor otherwise soluble proteins in the outer cell membrane. Lpp serves to direct the fusion protein, while a unstructured linker (GGGSGGGS) serves to separate OmpA from its cargo in order to avoid interactions between the two proteins.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">The expressed protein is structurally identical to BBa_K103006. This version of the biobrick has been moved into the new backbone, pSB1C3, which contains the gene for chloramphenicol resistance. This resistance gene contains an internal SacI site, making the SacI site at the end of the gene almost unusable for RE cloning. BBa_K1489003 mutagenizes this SacI site into a KasI in order to avoid this issue.</p><br />
<p><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;"> </span></p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="line-height: 12.0000009536743px; color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; text-decoration: underline;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">Part:BBa_K1489000 | </span></span><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap; text-decoration: underline;">pBAD-RBS-Bovine Galectin 1-B0012 Terminator</span></strong></span></p><br />
<p><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/parts/4/42/PSB1C3_-_BBa_K1489000.png" border="0" width="515" height="335" style="border: 0px; display: block; margin-left: auto; margin-right: auto;" /></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;">A soluble his-tagged galectin from Bos taurus, expressed under Arabinose induction</span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px; text-align: center;"><span style="font-size: 20px; font-weight: bold;">pBAD-RBS-Bovine Galectin-1-B0012 Terminator</span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">Soluble beta-galactoside binding protein (galectin) originally from Bos taurus. It is believed to expressed both intracellularly and extracellularly, where they serve to binding various types of beta-galactosides and initialize signal cascades. It is believed to be a dimer in its active form, as is true with most mammalian galectins.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">This biobrick is under the pBAD promoter (BBa_K206000) with attached RBS (BBa_B0034), and utilizes an additional RNA polymerase terminator (BBa_B0012).</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">UMaryland 2014 is interested in utilizing this part to investigate whether E. coli can recognize and bind to surface carbohydrates on a marine pathogen. Other uses of this part lie in the recognition of various carbohydrate ligands and potential activation of signal transduction pathways.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;"> </p><br />
<p style="text-align: center;"> </p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="text-align: left; line-height: 17.9400005340576px; white-space: pre-wrap;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; text-decoration: underline;">Part:BBa_K1489004 | </span></span><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; line-height: 17.9400005340576px; white-space: pre-wrap; text-decoration: underline;">pBAD-RBS-Lpp-OmpA-Bovine Galectin 1-B0012 Terminator</span></strong></span></p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; line-height: 17.9400005340576px; white-space: pre-wrap; text-decoration: underline;"><img src="https://static.igem.org/mediawiki/parts/0/0f/PSB1C3_-_BBa_K1489004.png" border="0" width="515" height="335" /></span></strong></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">His-tagged bovine galectin 1 attached to the c-terminus of OmpA, expressed under arabinose induction</span></span></p><br />
<p> </p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 20px; font-weight: bold; line-height: 19.5px; text-align: start;">pBAD-RBS-OmpA-Bovine Galectin-1-B0012 Terminator</span></span></p><br />
<p style="text-align: left;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 13px; line-height: 19.5px; text-align: start;">A fusion protein of OmpA (BBa_K1489002) fused to Bovine Galectin-1 (BBa_K1489001). Used to bind sugars at the outer cell membrane. Under pBAD regulation.</span></span></p><br />
<p style="text-align: left;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"> </span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"><span style="text-decoration: underline;"><strong>Part:BBa_K1489003 | Revised Lpp-OmpA</strong></span></span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"><span style="text-decoration: underline;"><strong><img src="https://static.igem.org/mediawiki/parts/9/90/PSB1C3_-_BBa_K1489003.png" border="0" width="515" height="335" /></strong></span></span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"> Biobrick 1, with a SacI site at the C-terminus replaced by a KasI site. This improvement was made in order to re-enable RE cloning; a SacI site exists in the chloramphenicol resistance gene.</span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 20px; font-weight: bold; line-height: 19.5px; text-align: start;">Lpp-OmpA-Linker with KasI site</span></span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">Outer membrane protein A (OmpA) is a common outer membrane protein found in bacteria that serves various purposes. For our purposes, the beta-barrel transmembrane domain can be utilized to anchor otherwise soluble proteins in the outer cell membrane. Lpp serves to direct the fusion protein, while a unstructured linker (GGGSGGGS) serves to separate OmpA from its cargo in order to avoid interactions between the two proteins.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">The expressed protein is almost structurally identical to BBa_K103006, with the terminal serine on the linker replaced by an alanine. This change was made in order to convert the previous SacI site (GAGCTC), which was unusable for RE cloning due to a second SacI site being present in the plasmid backbone, into a KasI site (GGCGCC). The KasI site codes for Gly-Ala, while the SacI site codes for Gly-Ser. This amino acid substitution is unlikely to impact the function of Lpp-OmpA-Linker in a major way.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;"> </p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;"> </p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">Part:BBa_K1489001 | Bovine Galectin-1</span></span></strong></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/parts/4/4c/PSB1C3_-_BBa_K1489001.png" border="0" width="515" height="335" style="border: 0;" /></span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">This is the Bovine Galectin which should interact with beta-galactosides and intialize a signal cascase</span></span></p><br />
<p style="text-align: center;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 20px; font-weight: bold; line-height: 19.5px; text-align: start;">Bovine Galectin-1</span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">Soluble beta-galactoside binding protein (galectin) originally from Bos taurus. It is believed to expressed both intracellularly and extracellularly, where they serve to binding various types of beta-galactosides and initialize signal cascades. It is believed to be a dimer in its active form, as is true with most mammalian galectins.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">UMaryland 2014 is interested in utilizing this part to investigate whether E. coli can recognize and bind to surface carbohydrates on a marine pathogen. Other uses of this part lie in the recognition of various carbohydrate ligands and potential activation of signal transduction pathways.</p> <br />
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<div class="yjsg7_before"><div id="yjsg7" class="yjsg_grid yjsgsitew"><div id="user23" class="yjsgxhtml only_mod"><div class="yjsquare modid117"><div class="h2_holder"><h2 class="module_title"><span class="title_split titlesplit0">About</span> <span class="title_split titlesplit1">Umaryland</span></h2></div><div class="yjsquare_in"><p>UMaryland 2014 is the inaugural iGEM team of the University of Maryland, College Park. We are a combined effort of several departments and numerous faculty mentors. Although it is only our first year, we believe our hard work and dedication has paid off. We can't wait for this year's competition! GO TERPS!</p></div></div></div></div></div> <div class="footer_holders footer"><!-- footer --><br />
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<option value="https://2014.igem.org/Team:UMaryland">&nbsp;Homepage</option><br />
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<option value="https://2014.igem.org/Team:UMaryland/meetourteam">&nbsp;--Meet Our Team Members</option><br />
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<option value="https://2014.igem.org/Team:UMaryland/project/futureapplications">&nbsp;--future applications</option><br />
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<option value="https://2014.igem.org/Team:UMaryland/notebook">&nbsp;--Wetlab Notebook</option><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/project/background
Team:UMaryland/project/background
2014-10-17T19:41:30Z
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<h1 class="article_title"><br />
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Background-Motivation <br />
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<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: center;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"><em><span style="text-decoration: underline;"><strong>Background</strong></span></em><br /></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">We at UMaryland iGEM are developing a biosensor to detect for the presence of an oyster pathogen, </span><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">Perkinsus marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">. This parasite is commonly found over the eastern shore of North America, including in Maryland’s own Chesapeake Bay. </span><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">, a unicellular protist, causes an oyster infection known as Dermo (Espinosa, Winnicki, &amp; Allam, 2013). The pathogen has been shown to infest oysters by entering their hemocytes via induced phagocytosis. When inside the hemocyte, the parasite is capable of resisting the reactive oxygen species generated by the oyster immune system and modulating the apoptosis of infected hemocyte cells to spread itself further (Espinosa et al., 2013). This leads to proliferation inside the oyster hemocyte, eventually lysing the host cell and releasing more </span><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"> (Allam et al., 2013). This process leads to massive tissue damage and, ultimately, the death of the oyster.</span></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">It is believed that </span><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">P. marinus </span><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">enters utilizing a galectin, a galactose-binding protein, on the hemocyte cell surface, CvGal1, which allows it to bind preferentially to </span><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"> (Schott, Pecher, Okafor, &amp; Vasta, 2003). Our aim is to exploit this property by expressing the galectin on the surface of </span><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">E. coli</span><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"> bacterial cells by attaching it to the C-terminus of an OmpA transmembrane protein. These bacteria will thus bind to any </span><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"> present in a sample of water. Our long-term goal is to incorporate these biosensors into a lab-on-a-chip device, requiring only small samples and allowing quick response times for detecting this pathogen.</span></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">This year, we have focused on working with the bovine galectin-1 in place of CvGal1. At only 14 kDa in size, the bovine galectin was chosen as a simpler candidate for expression on the surface of the membrane (Liao, Kapadia, Ahmed, Vasta, &amp; Herzberg, 1994). Furthermore, this bovine galectin-1 was one of three galectins used for the homology modeling of CvGal1 due to their similar binding domains (Tasumi &amp; Vasta, 2007). Though bovine galectin-1 is a two-domain galectin, compared to the four domains of CvGal1 (Feng et al., 2013), it is believed that the bovine galectin has potential to bind the same ligand present on </span><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"> that CvGal1 binds. Additionally, we have optimized the bovine galectin-1 gene, most notably for better protein folding and BioBrick friendliness.</span></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">Although laboratory methods for Dermo detection using PCR exist, they are currently not capable of detecting the disease in real time. In the time needed to conduct a laboratory test, Dermo will have easily spread to new locations. If we can pinpoint the location of Dermo on-site, then we can take action ultimately to eliminate the parasite from eastern oyster habitats.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">References</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">Allam, B., Carden, W. E., Ward, J. E., Ralph, G., Winnicki, S., &amp; Espinosa, E. P. (2013). Early host-pathogen interactions in marine bivalves: Evidence that the alveolate parasite Perkinsus marinus infects through the oyster mantle during rejection of pseudofeces. </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">Journal of Invertebrate Pathology</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">, </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">113</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">(1), 26-34.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">Espinosa, E. P., Winnicki, S., &amp; Allam, B. (2013). Early host–pathogen interactions in a marine bivalve: Crassostrea virginica pallial mucus modulates Perkinsus marinus growth and virulence. </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">Diseases of Aquatic Organisms</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">, </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">104</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">(3), 237-247.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">Feng, C., Ghosh, A., Amin, M. N., Giomarelli, B., Shridhar, S., Banerjee, A., ... &amp; Vasta, G. R. (2013). The galectin CvGal1 from the eastern oyster (Crassostrea virginica) binds to blood group A oligosaccharides on the hemocyte surface. </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">Journal of Biological Chemistry</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">, </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">288</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">(34), 24394-24409.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">Liao, D. I., Kapadia, G., Ahmed, H., Vasta, G. R., &amp; Herzberg, O. (1994). Structure of S-lectin, a developmentally regulated vertebrate beta-galactoside-binding protein. </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">Proceedings of the National Academy of Sciences</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">, </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">91</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">(4), 1428-1432.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">Tasumi, S., &amp; Vasta, G. R. (2007). A galectin of unique domain organization from hemocytes of the Eastern oyster (Crassostrea virginica) is a receptor for the protistan parasite Perkinsus marinus. </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">The Journal of Immunology</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">, </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">179</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">(5), 3086-3098.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">Schott, E. J., Pecher, W. T., Okafor, F., &amp; Vasta, G. R. (2003). The protistan parasite Perkinsus marinus is resistant to selected reactive oxygen species. </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">Experimental Parasitology</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">, </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">105</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">(3-4), 232-240.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"> </p><br />
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<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicschool">&nbsp;--Public School Outreach</option><br />
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<option value="https://2014.igem.org/Team:UMaryland/humanprac/community">&nbsp;--Community Interaction</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicawareness">&nbsp;--Public Awareness</option><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/project/overview
Team:UMaryland/project/overview
2014-10-17T19:41:04Z
<p>Nbb826: </p>
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Overview <br />
</a><br />
</h1><br />
<br />
<br />
<br />
<p style="text-align: left;"> </p><br />
<p style="margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: center;" dir="ltr"><em><span style="text-decoration: underline;"><strong><span style="vertical-align: baseline;"><span style="color: #000000; font-family: Arial;"><span style="font-size: 15.5555562973022px; line-height: 19.1666679382324px; white-space: pre-wrap;">Overview</span></span></span></strong></span></em></p><br />
<p style="margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: center;" dir="ltr"><em><span style="text-decoration: underline;"><strong><span style="vertical-align: baseline;"><span style="color: #000000; font-family: Arial;"><span style="font-size: 15.5555562973022px; line-height: 19.1666679382324px; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/parts/f/f2/Umarylandigeminfographic.jpg" border="0" width="402" height="256" /></span></span></span></strong></span></em></p><br />
<p style="margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: left;" dir="ltr"><span style="vertical-align: baseline;"><span style="color: #000000; font-family: Arial;"><span style="font-size: 15.5555562973022px; line-height: 19.1666679382324px; white-space: pre-wrap;">The UMaryland iGem team opted for a project that hits close to home; with the Chesapeake Bay in our backyard, we have the potential to make a direct impact on the community. As Maryland’s most important natural landmark, the Chesapeake Bay is an integral part of our state’s identity. The bay is home to a diverse ecosystem, and is an economic boon to the area, supporting both the seafood industry and local tourism. Since the health of the bay is important, we looked into current problems in need of attention. In particular, the predicament of the Chesapeake’s oysters piqued our interest. As oyster populations declined over the years, the absence of this filtration system left a host of problems, including decreased water clarity and rising chemical pollutant levels. If the oyster populations can be maintained, it will improve the overall health of the Chesapeake. One way to combat oyster fatalities is to develop a way to monitor Dermo infestations. </span></span> </span></p><br />
<p style="text-align: left;"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: center;" dir="ltr"> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: center;" dir="ltr"><em><strong><span style="text-decoration: underline;"><span style="font-size: 15px; font-family: Arial; color: #000000; font-variant: normal; text-decoration: underline; vertical-align: baseline; white-space: pre-wrap; background-color: #ffffff;">Applications in the lab</span></span></strong></em></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">It is believed that </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">P. marinus </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">enters oyster hemocytes utilizing a galectin on the cell’s surface, CvGal1, that allows binds preferentially to </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">. Our aim is to exploit CvGal1’s ability to bind to </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> in order to design a biosensor for the parasite. By expressing the galectin on the surface of </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">E. coli</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> and triggering a signal transduction cascade within the bacterial cell, the </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">E. coli</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> could produce Green Fluorescent Protein. Thus, </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">P. marinus </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">binding to CvGal1 would trigger the expression of GFP within </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">E. coli, </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> creating a visual marker for the parasite. The majority of this project will be accomplished through genetic cloning and bacterial transformation. Although laboratory methods for Dermo detection using Polymerase Chain Reaction exist, they are currently not capable of detecting the disease in real time. In the time needed to conduct a laboratory test, Dermo could have easily spread to new locations. This biosensor would help to accurately pinpoint Dermo infestation levels in real time. If we can pinpoint the location of Dermo, as elucidated in this research proposal, then we can ultimately develop a biosensor that can react to Dermo with targeted destruction. </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"><strong id="docs-internal-guid-809a97bc-f5c8-6033-4262-354a17c98c1d" style="font-weight: normal;"><br /><br /></strong></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: center;" dir="ltr"><span style="text-decoration: underline;"><strong><em><span style="font-size: 15px; font-family: Arial; color: #000000; font-variant: normal; text-decoration: underline; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">Brief overview of servicing the community</span></em></strong></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">We having been traveling around Maryland conducting various community outreach efforts by visiting local schools and oyster farms. During each visit, members present our project and spread awareness on the current state of the Chesapeake Bay, as well as ways individuals can help service the Bay. Some presentations are made on synthetic biology to increase student participation and interest in the sciences. We also have a committee dedicated to creating an educational computer game that teaches others about the issues that the Chesapeake Bay currently faces. It is our team’s hope that our research results to be presented at the iGEM jamboree will have a profound and positive impact on the health of the Chesapeake Bay and oyster populations worldwide</span></p><br />
<p><span><span> </span></span></p><br />
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<li class="item-101 lifirst "><a class="afirst" href="https://2014.igem.org/Team:UMaryland" >Homepage</a></li><li class="item-104"><a href="https://2014.igem.org/Team:UMaryland/photos" >Photos</a></li><li class="item-105 parent"><a href="https://2014.igem.org/Team:UMaryland/meetourteam" >Team</a><ul class="yjsgacc-dropdown" id="modid19-menu105"><li class="item-293"><a href="https://2014.igem.org/Team:UMaryland/meetourteam" >Meet Our Team Members</a></li><li class="item-116"><a href="https://igem.org/Team.cgi?id=1489" >iGEM Page</a></li><li class="item-117"><a href="https://2014.igem.org/Team:UMaryland/team/attributions" >Attributions</a></li><li class="item-300"><a href="https://2014.igem.org/Team:UMaryland/team/sponsors" >Sponsors</a></li></ul></li><li class="item-112 active parent"><a class="yjscroll" href="https://2014.igem.org/Team:UMaryland/project/overview" >Project</a><ul class="yjsgacc-dropdown" id="modid19-menu112"><li class="item-138 current-side active "><a class=" isactivea" href="https://2014.igem.org/Team:UMaryland/project/overview" >Overview</a></li><li class="item-145"><a href="https://2014.igem.org/Team:UMaryland/project/background" >Background</a></li><li class="item-144"><a href="https://2014.igem.org/Team:UMaryland/project/biobricks" >parts (BioBricks)</a></li><li class="item-102"><a href="https://2014.igem.org/Team:UMaryland/project/results" target="_blank" >Results</a></li><li class="item-254"><a href="https://2014.igem.org/Team:UMaryland/project/futureapplications" >future applications</a></li><li class="item-301"><a href="https://igem.org/Safety/Safety_Form?team_id=1489" >Safety</a></li></ul></li><li class="item-234 parent"><a href="https://2014.igem.org/Team:UMaryland/notebook" >Notebook</a><ul class="yjsgacc-dropdown" id="modid19-menu234"><li class="item-303"><a href="https://2014.igem.org/Team:UMaryland/notebook" >Wetlab Notebook</a></li></ul></li><li class="item-123 parent"><a href="https://2014.igem.org/Team:UMaryland/humanprac/overview" >Human Practices</a><ul class="yjsgacc-dropdown" id="modid19-menu123"><li class="item-106"><a href="https://2014.igem.org/Team:UMaryland/humanprac/overview" >Overview | Mission Statement</a></li><li class="item-129"><a href="https://2014.igem.org/Team:UMaryland/humanprac/publicschool" >Public School Outreach</a></li><li class="item-131"><a href="https://2014.igem.org/Team:UMaryland/humanprac/community" >Community Interaction</a></li><li class="item-132 lilast"><a class="alast" href="https://2014.igem.org/Team:UMaryland/humanprac/publicawareness" >Public Awareness</a></li></ul></li></ul></div></div><br />
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<option value="https://2014.igem.org/Team:UMaryland">&nbsp;Homepage</option><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/team/attributions
Team:UMaryland/team/attributions
2014-10-17T19:40:34Z
<p>Nbb826: </p>
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href="https://2014.igem.org/Team:UMaryland/project/futureapplications"><span class="yjm_has_none"><span class="yjm_title">future applications</span></span></a></span></li><li class="item301 level1 lilast"><span class="mymarg"><a class="yjanchor last" href="https://igem.org/Safety/Safety_Form?team_id=1489"><span class="yjm_has_none"><span class="yjm_title">Safety</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li><li class="haschild item234 level0"><span class="child"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/notebook"><span class="yjm_has_none"><span class="yjm_title">Notebook</span></span></a></span><ul class="subul_main level1"><li class="bl"></li><li class="tl"></li><li class="tr"></li><li class="item303 level1 first lilast"><span class="mymarg"><a class="yjanchor firstlast" href="https://2014.igem.org/Team:UMaryland/notebook"><span class="yjm_has_none"><span class="yjm_title">Wetlab Notebook</span></span></a></span></li><li 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Attributions <br />
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<h1 style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">We Would like to thank all who have helped make this year possible!</span></h1><br />
<h2 style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> <br />A Special Thanks to the following contributors: </span></h2><br />
<p><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/parts/0/0e/Bentley.jpg" border="0" width="155" height="232" style="border: 0;" /></span></p><br />
<h3 style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">Dr. William Bentley</span></strong></h3><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="color: #000000; font-family: Arial; font-size: 15px; font-weight: normal; line-height: 1.15; text-indent: 36pt; white-space: pre-wrap; background-color: transparent;"> Distinguished professor and chair, Fischell Department of Bioengineering University of Maryland</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="color: #000000; font-family: Arial; font-size: 15px; font-weight: normal; line-height: 1.15; text-indent: 36pt; white-space: pre-wrap; background-color: transparent;">Dr. Bentley was helpful talking through the initial stages of the project, representing the schools Bioengineering department and getting out name out to campus, as well as a major supporter of the project, in addition to providing lab instruments, resources, and offering cellular constructs.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="color: #000000; font-family: Arial; font-size: 15px; font-weight: normal; line-height: 1.15; text-indent: 36pt; white-space: pre-wrap; background-color: transparent;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="color: #000000; font-family: Arial; font-size: 15px; font-weight: normal; line-height: 1.15; text-indent: 36pt; white-space: pre-wrap; background-color: transparent;"><img src="https://static.igem.org/mediawiki/parts/a/a6/JohnBuchner.jpg" border="0" width="126" height="188" style="border: 0;" /></span></p><br />
<h3 style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">Dr. John Buchner</span></strong></h3><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Lecturer, Department of Cell Biology and Molecular Genetics University of Maryland.</span><span style="line-height: 12.0000009536743px;">John Buchner is lecturer and lab coordinator for general microbiology, and a lab coordinator for microbial genetics. His microbiology class is in I Series courses at UMD. He is a member of the HPI group, a faculty collaboration intent improving the educational experience of undergraduates, and publishing the findings. John is also a Chesapeake Fellow, and is dedicated raising awareness of sustainability to the students he teaches, and reducing material consumed in his classes.</span></p><br />
<p class="MsoNormal"> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Dr. Buchner was instrumental in helping our students design the project, providing many insights into how proteins function through the cell membranes, as well as signal transduction pathways.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/parts/4/48/BenWoodard.jpg" border="0" width="145" height="239" /></span></p><br />
<h3 style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">Dr. Ben Woodard</span></strong></h3><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Director, Biotechnology Research and Education Program</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Director, Bioprocess Scale-up Facility</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Dr. Woodard was a valuable research throughout the planning phase of the project, providing insight on the potential commercial aspect of the product and keeping our team on track with realistic expectations.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<h3 style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">John Wilhelm III</span></strong></h3><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="color: #000000; font-family: Arial; font-size: 15px; line-height: 17.25px; white-space: pre-wrap;">Regional Sales manager at Integrated DNA technologies</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="color: #000000; font-family: Arial; font-size: 15px; line-height: 17.25px; white-space: pre-wrap;">Mr. Wilhelm provided a wealth of knowledge regarding the creation of our constructs and the different options availible, as well as introducing us to synthetic gene construction. IDT was responsible for creating our G-Blocks used for CVGAL1 and Bovine Galectin-1</span></p><br />
<p><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span><img src="https://static.igem.org/mediawiki/parts/8/8a/Vastaheadshot.jpg" border="0" alt="Gerardo R Vasta " width="175" height="219" style="border: 0;" /></p><br />
<h3 style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><strong>Dr. Gerardo Vasta</strong></span></h3><br />
<p><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Professor, Department of Microbiology and Immunology University of Maryland School of Medicine</span></p><br />
<p><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Dr. Vasta was extremely helpful with our questions regarding Dermo, specifically the ligand CVGAL1 and the interaction between Dermo and E.Coli as well providing strategies for expressing and optimizing this protein.</span></p><br />
<p><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/parts/1/15/Herzberg.jpg" border="0" width="180" height="190" style="border: 0;" /></span></p><br />
<h3 style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><strong>Dr. Osnat Herzberg</strong></span></h3><br />
<p><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Professor Department of Chemistry and Biochemistry University of Maryland at Shady Grove</span></p><br />
<p><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Dr. Herzberg contributed to our protein design, offering to check our sequences and provided insights into the bovine-galectin that we created.</span></p><br />
<h2><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">A Special Thanks to the Following Departments/Laboratories:</span></h2><br />
<h3 style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">The Cell Biology and Molecular Genetics</span></strong></h3><br />
<p><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">A special thanks for Cell Biology and Molecular Genetics for offering us a lab space throughout the duratation of the summer in which we conducted our research.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<h3 style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">Dr. Kahn's Laboratory</span></strong></h3><br />
<p><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;"><br /></span></strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">A special thanks to The Kahn Lab group for offering us lab space throughout the fall semester, providing additional materials we needed for research and offering storage space. Additionally Dr. Kahn and Dr. Quimby performed the ran the final western blot which was included in our results</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">Dr. Bentley's Laboratory</span></strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong style="line-height: 1.15;"><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;"><br /></span></strong><span style="color: #000000; font-family: Arial;"><span style="font-size: 15px; line-height: 17.25px; white-space: pre-wrap;">A special thanks to The Bentley Lab group for permitting us the use of their facilities,</span></span><span style="color: #000000; font-family: Arial; font-size: 15px; line-height: 17.25px; white-space: pre-wrap;">including multiple analysis machines without which we could not have completed our research,</span><span style="font-size: 15px; line-height: 17.25px; white-space: pre-wrap; color: #000000; font-family: Arial; text-indent: 36pt;"> providing enzymes and other research materials.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; line-height: 17.25px; white-space: pre-wrap; color: #000000; font-family: Arial; text-indent: 36pt;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">Dr. Eisensteins Laboratory</span></strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;"><br /></span></strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">A special thanks to the Eisenstein Lab group for providing us with research materials, as well as allowing us to run experiments at their location</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;"> </span></p><br />
<h3 style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">Fischell Department of Bioengineering</span></strong></h3><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">A. James Clark School of Engineering</span></strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">College of Computer, Math, and Natural Sciences</span></strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;"><span style="line-height: 1.15; text-indent: 36pt; background-color: transparent;">Integrated Life Sciences (ILS)</span></span></strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"> </p><br />
<h3><strong><span id="docs-internal-guid-809a97bc-c77f-0ff5-d846-9d15e5944245"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;"> National Scholarships Office (Leadership and Discovery Grant)</span></span></strong></h3> </div><br />
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Team:UMaryland/meetourteam
2014-10-17T19:40:00Z
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<h1>Meet our awesome team</h1><br />
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<p>Find our more about the three groups of contributors that made our project possible</p></div></div></div></div> </div><br />
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<a href="https://2014.igem.org/Team:UMaryland/meetourteam/tanya"><br />
Tanya Gordonov </a><br />
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<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/1/1d/TanyaGordonov.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
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<p style="text-align: center;"><strong>Graduate Advisor | Bioengineering</strong></p><br />
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<a href="https://2014.igem.org/Team:UMaryland/meetourteam/adip"><br />
Adip Bhargav </a><br />
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<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/1/13/AdipBhargav.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate | Junior | Neurobiology &amp; Physiology | French Minor</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
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<a href="https://2014.igem.org/Team:UMaryland/meetourteam/aditya"><br />
Aditya Biswas </a><br />
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<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/6/6d/Adityabiswas.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
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<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate | Junior | Bioengineering</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
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<a href="https://2014.igem.org/Team:UMaryland/meetourteam/navadeep"><br />
Navadeep Boruah </a><br />
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<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/0/08/Navadeep2.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
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<p style="text-align: center;"><strong>Graduate Advisor | Chemical and Biomolecular Engineering</strong></p><br />
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<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/navadeep"><br />
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<a href="https://2014.igem.org/Team:UMaryland/meetourteam/andrew"><br />
Andrew Zhao </a><br />
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<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/0/06/AndrewZhao.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
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<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate | Sophomore | Bioengineering</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
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<a href="https://2014.igem.org/Team:UMaryland/meetourteam/kevin"><br />
Kevin Pineault </a><br />
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<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/4/42/KevinPineault.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
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<p style="text-align: center;"><strong>Undergraduate | Junior | Bioengineering</strong></p><br />
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<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/kevin"><br />
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<a href="https://2014.igem.org/Team:UMaryland/meetourteam/iowis"><br />
Iowis Zhu </a><br />
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<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/d/d1/IowisZhu.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
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<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate | Junior | Biochemistry/Cell Biology and Molecular Genetics</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
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<a href="https://2014.igem.org/Team:UMaryland/meetourteam/chris"><br />
Chris Euerle </a><br />
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<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/d/d6/Chriseuerle.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
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<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate | Junior | Computer Science and Neurobiology &amp; Physiology</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
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<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/chris"><br />
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<a href="https://2014.igem.org/Team:UMaryland/meetourteam/boots"><br />
Dr. Boots Quimby </a><br />
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<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/f/f0/Bootsquimby.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
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<p style="text-align: center;"><strong>Faculty Advisor | Integrated Life Sciences</strong></p><br />
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Janine Taira </a><br />
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<div class="clr"></div><br />
<div class="newsitem_text"><br />
<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/4/49/JanineTaira.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate | Sophomore | Neurobiology &amp; Physiology</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
<p> </p><br />
</div><br />
<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/janine"><br />
<span><br />
Read more: Janine Taira </span><br />
</a><br />
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<span class="article_separator">&nbsp;</span><br />
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<div class="float-left width33"><br />
<br />
<div class="news_item_c teamPage"><br />
<h2 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/meetourteam/camila"><br />
Camila Uechi </a><br />
</h2><br />
<div class="clr"></div><br />
<div class="newsitem_text"><br />
<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/6/63/Camilauechi.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate | Junior | Bioengineering</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
<p> </p><br />
</div><br />
<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/camila"><br />
<span><br />
Read more: Camila Uechi </span><br />
</a><br />
</div><br />
<span class="article_separator">&nbsp;</span><br />
</div><br />
<br />
<br />
<div class="last float-left width33"><br />
<br />
<div class="news_item_c teamPage"><br />
<h2 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/meetourteam/jason"><br />
Jason Kahn </a><br />
</h2><br />
<div class="clr"></div><br />
<div class="newsitem_text"><br />
<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/d/d1/JasonKahn.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong>Faculty Advisor|Biochemistry</strong></p><br />
<p> </p><br />
</div><br />
<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/jason"><br />
<span><br />
Read more: Jason Kahn </span><br />
</a><br />
</div><br />
<span class="article_separator">&nbsp;</span><br />
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<span class="row-separator"></span><br />
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<div class="first float-left width33"><br />
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<div class="news_item_c teamPage"><br />
<h2 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/meetourteam/nathan"><br />
Nathan Barber </a><br />
</h2><br />
<div class="clr"></div><br />
<div class="newsitem_text"><br />
<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/8/81/NathanBarber.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate | Senior | Bioengineering | Engineering Leadership Minor</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
<p> </p><br />
</div><br />
<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/nathan"><br />
<span><br />
Read more: Nathan Barber </span><br />
</a><br />
</div><br />
<span class="article_separator">&nbsp;</span><br />
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<br />
<div class="float-left width33"><br />
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<div class="news_item_c teamPage"><br />
<h2 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/meetourteam/pavel"><br />
Pavel Vlasov </a><br />
</h2><br />
<div class="clr"></div><br />
<div class="newsitem_text"><br />
<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/5/55/Pavelvlasov.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate | Sophomore | Biochemistry/Cell Biology &amp; Molecular Genetics</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
<p> </p><br />
</div><br />
<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/pavel"><br />
<span><br />
Read more: Pavel Vlasov </span><br />
</a><br />
</div><br />
<span class="article_separator">&nbsp;</span><br />
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<br />
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<div class="last float-left width33"><br />
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<div class="news_item_c teamPage"><br />
<h2 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/meetourteam/ed"><br />
Dr. Edward Eisenstein </a><br />
</h2><br />
<div class="clr"></div><br />
<div class="newsitem_text"><br />
<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/7/77/Edeisenstein.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong>Faculty Advisor | Bioengineering</strong></p><br />
<p> </p><br />
</div><br />
<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/ed"><br />
<span><br />
Read more: Dr. Edward Eisenstein </span><br />
</a><br />
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<li class="first"><br />
<span class="catitem_title"><a href="https://2014.igem.org/Team:UMaryland/meetourteam/grad"><br />
Graduate Student Advisors</a><br />
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<br />
<br />
<br />
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<span class="catitem_title"><a href="https://2014.igem.org/Team:UMaryland/meetourteam/undergrad"><br />
Undergraduate Students</a><br />
</span><br />
<br />
<br />
<br />
</li><br />
<li class="last"><br />
<span class="catitem_title"><a href="https://2014.igem.org/Team:UMaryland/meetourteam/faculty"><br />
Faculty Advisors</a><br />
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<div class="yjsg7_before"><div id="yjsg7" class="yjsg_grid yjsgsitew"><div id="user23" class="yjsgxhtml only_mod"><div class="yjsquare modid117"><div class="h2_holder"><h2 class="module_title"><span class="title_split titlesplit0">About</span> <span class="title_split titlesplit1">Umaryland</span></h2></div><div class="yjsquare_in"><p>UMaryland 2014 is the inaugural iGEM team of the University of Maryland, College Park. We are a combined effort of several departments and numerous faculty mentors. Although it is only our first year, we believe our hard work and dedication has paid off. We can't wait for this year's competition! GO TERPS!</p></div></div></div></div></div> <div class="footer_holders footer"><!-- footer --><br />
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<!-- end footer --><br />
<script type="text/javascript"> var logo_w = '240'; var site_w = '1200'; var site_f = '13px'; var sp='https://2014.igem.org/Team:UMaryland'; var tp ='baseline'; var compileme =0; var fontc ='baseline_73331408039633'; var bootstrapv='bootstrap2'; var yver='2'; var yjsglegacy='1'; var yjsgrtl='2'; var menuanimation='fade';var menuanimationspeed=300; var YJSG_topmenu_font = '13px'; (function($){ $(window).load(function(){ $('.horiznav').SmoothDropJQ({ contpoz:0, horizLeftOffset: 20, horizRightOffset: -20, horizTopOffset: 20, verticalTopOffset:30, verticalLeftOffset: 10, maxOutside: 50 }); }) })(jQuery); </script><br />
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<span class="yjmm_select" id="yjmm_selectid">Meet Our Team Members</span><br />
<select id="mmenu" class="yjstyled"><br />
<option value="https://2014.igem.org/Team:UMaryland">&nbsp;Homepage</option><br />
<option value="https://2014.igem.org/Team:UMaryland/photos">&nbsp;Photos</option><br />
<option value="https://2014.igem.org/Team:UMaryland/meetourteam">&nbsp;Team</option><br />
<option value="https://2014.igem.org/Team:UMaryland/meetourteam" selected="selected">&nbsp;--Meet Our Team Members</option><br />
<option value="https://igem.org/Team.cgi?id=1489">&nbsp;--iGEM Page</option><br />
<option value="https://2014.igem.org/Team:UMaryland/team/attributions">&nbsp;--Attributions</option><br />
<option value="https://2014.igem.org/Team:UMaryland/team/sponsors">&nbsp;--Sponsors</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/overview">&nbsp;Project</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/overview">&nbsp;--Overview</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/background">&nbsp;--Background</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/biobricks">&nbsp;--parts (BioBricks)</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/results">&nbsp;--Results</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/futureapplications">&nbsp;--future applications</option><br />
<option value="https://igem.org/Safety/Safety_Form?team_id=1489">&nbsp;--Safety</option><br />
<option value="https://2014.igem.org/Team:UMaryland/notebook">&nbsp;Notebook</option><br />
<option value="https://2014.igem.org/Team:UMaryland/notebook">&nbsp;--Wetlab Notebook</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/overview">&nbsp;Human Practices</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/overview">&nbsp;--Overview | Mission Statement</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicschool">&nbsp;--Public School Outreach</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicschool">&nbsp;---HighSchool</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/community">&nbsp;--Community Interaction</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicawareness">&nbsp;--Public Awareness</option><br />
</select><br />
</div> </body><br />
</html></div>
Nbb826
http://2014.igem.org/Team:UMaryland
Team:UMaryland
2014-10-17T19:39:03Z
<p>Nbb826: </p>
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<li id="current" class=" active item101 level0 first"><span class="mymarg"><a class="yjanchor first activepath " href="https://2014.igem.org/Team:UMaryland"><span class="yjm_has_none"><span class="yjm_title">Homepage</span></span></a></span></li><li class="item104 level0"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/photos"><span class="yjm_has_none"><span class="yjm_title">Photos</span></span></a></span></li><li class="haschild item105 level0"><span class="child"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/meetourteam"><span class="yjm_has_none"><span class="yjm_title">Team</span></span></a></span><ul class="subul_main level1"><li class="bl"></li><li class="tl"></li><li class="tr"></li><li class="item293 level1 first"><span class="mymarg"><a class="yjanchor first" href="https://2014.igem.org/Team:UMaryland/meetourteam"><span class="yjm_has_none"><span class="yjm_title">Meet Our Team Members</span></span></a></span></li><li class="item116 level1"><span class="mymarg"><a class="yjanchor " href="https://igem.org/Team.cgi?id=1489"><span class="yjm_has_none"><span class="yjm_title">iGEM Page</span></span></a></span></li><li class="item117 level1"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/team/attributions"><span class="yjm_has_none"><span class="yjm_title">Attributions</span></span></a></span></li><li class="item300 level1 lilast"><span class="mymarg"><a class="yjanchor last" href="https://2014.igem.org/Team:UMaryland/team/sponsors"><span class="yjm_has_none"><span class="yjm_title">Sponsors</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li><li class="haschild item112 level0"><span class="child"><a class="yjanchor yjscroll" href="#"><span class="yjm_has_none"><span class="yjm_title">Project</span></span></a></span><ul class="subul_main level1"><li class="bl"></li><li class="tl"></li><li class="tr"></li><li class="item138 level1 first"><span class="mymarg"><a class="yjanchor first" href="https://2014.igem.org/Team:UMaryland/project/overview"><span class="yjm_has_none"><span class="yjm_title">Overview</span></span></a></span></li><li class="item145 level1"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/project/background"><span class="yjm_has_none"><span class="yjm_title">Background</span></span></a></span></li><li class="item144 level1"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/project/biobricks"><span class="yjm_has_none"><span class="yjm_title">parts (BioBricks)</span></span></a></span></li><li class="item102 level1"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/project/results" target="_blank" ><span class="yjm_has_none"><span class="yjm_title">Results</span></span></a></li><li class="item254 level1"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/project/futureapplications"><span class="yjm_has_none"><span class="yjm_title">future applications</span></span></a></span></li><li class="item301 level1 lilast"><span class="mymarg"><a class="yjanchor last" href="https://igem.org/Safety/Safety_Form?team_id=1489"><span class="yjm_has_none"><span class="yjm_title">Safety</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li><li class="haschild item234 level0"><span class="child"><a class="yjanchor " href="#"><span class="yjm_has_none"><span class="yjm_title">Notebook</span></span></a></span><ul class="subul_main level1"><li class="bl"></li><li class="tl"></li><li class="tr"></li><li class="item303 level1 first lilast"><span class="mymarg"><a class="yjanchor firstlast" href="https://2014.igem.org/Team:UMaryland/notebook"><span class="yjm_has_none"><span class="yjm_title">Wetlab Notebook</span></span></a></span></li><li class="right"></li><li 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href="https://2014.igem.org/Team:UMaryland/humanprac/publicschool"><span class="yjm_has_none"><span class="yjm_title">HighSchool</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li><li class="item131 level1"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/humanprac/community"><span class="yjm_has_none"><span class="yjm_title">Community Interaction</span></span></a></span></li><li class="item132 level1 lilast"><span class="mymarg"><a class="yjanchor last" href="https://2014.igem.org/Team:UMaryland/humanprac/publicawareness"><span class="yjm_has_none"><span class="yjm_title">Public Awareness</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li></ul></div><br />
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<p>Home of The University of Maryland's first iGEM team.</p><br />
<p><a href="https://2014.igem.org/Team:UMaryland/project/overview"><br /><br /></a></p><br />
<p><a href="https://2014.igem.org/Team:UMaryland/project/overview" target="_self"><img src="https://static.igem.org/mediawiki/parts/5/55/UMIgem_logo_%281%29.png" border="0" alt="Our research heavily focuses on saving the Chesapeake Bay through protecting the oysters" width="1000" height="364" onmouseover="this.src='https://static.igem.org/mediawiki/parts/f/f2/Umarylandigeminfographic.jpg'" onmouseout="this.src= 'https://static.igem.org/mediawiki/parts/5/55/UMIgem_logo_%281%29.png'" style="border: 0;" /></a></p></div></div></div></div> </div><br />
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<h4>Project</h4><br />
<p><img src="http://static.guim.co.uk/sys-images/Lifeandhealth/Pix/pictures/2008/11/05/oyster1.jpg" border="0" width="80" height="48" /></p><br />
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<h4>Notebook</h4><br />
<p><img src="https://static.igem.org/mediawiki/parts/c/c8/UmarylandNotebook.jpg" border="0" width="67" height="67" style="border: 0;" /></p><br />
<a class="rm_content" href="https://2014.igem.org/Team:UMaryland/notebook">Read More</a></div><br />
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<h4>human practices</h4><br />
<p><img src="https://static.igem.org/mediawiki/parts/9/97/UmarylandHumanpractices.png" border="0" width="99" height="72" style="border: 0;" /></p><br />
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<h4>Photo Album</h4><br />
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<div class="yjsg5_before"><div id="yjsg5" class="yjsg_grid yjsgsitew"><div id="user11" class="yjsgxhtml first_mod"><div class="yjsquare modid126"><div class="h2_holder"><h2 class="module_title"><span class="title_split titlesplit0">Meet</span> <span class="title_split titlesplit1">the</span> <span class="title_split titlesplit2">Team</span></h2></div><div class="yjsquare_in"><ul class="categories-module"><br />
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Graduate Student Advisors</a><br />
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Undergraduate Students</a><br />
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Faculty Advisors</a><br />
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<p><a href="https://2014.igem.org/Team:UMaryland/meetourteam" target="_self"><img src="https://static.igem.org/mediawiki/parts/7/7a/Umarylandlabphoto.jpg" border="0" alt="Our research heavily focuses on saving the Chesapeake Bay through protecting the oysters" width="500" height="150" onmouseover="this.src='https://static.igem.org/mediawiki/parts/d/d4/TeamPhoto.JPG'" onmouseout="this.src= 'https://static.igem.org/mediawiki/parts/7/7a/Umarylandlabphoto.jpg'" style="border: 0;" /></a></p></div></div></div></div></div> <div class="yjsg7_before"><div id="yjsg7" class="yjsg_grid yjsgsitew"><div id="user23" class="yjsgxhtml only_mod"><div class="yjsquare modid117"><div class="h2_holder"><h2 class="module_title"><span class="title_split titlesplit0">About</span> <span class="title_split titlesplit1">Umaryland</span></h2></div><div class="yjsquare_in"><p>UMaryland 2014 is the inaugural iGEM team of the University of Maryland, College Park. We are a combined effort of several departments and numerous faculty mentors. Although it is only our first year, we believe our hard work and dedication has paid off. We can't wait for this year's competition! GO TERPS!</p></div></div></div></div></div> <div class="footer_holders footer"><!-- footer --><br />
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<option value="https://2014.igem.org/Team:UMaryland/project/biobricks">&nbsp;--parts (BioBricks)</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/results">&nbsp;--Results</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/futureapplications">&nbsp;--future applications</option><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/project/futureapplications
Team:UMaryland/project/futureapplications
2014-10-17T19:30:30Z
<p>Nbb826: </p>
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<a href="https://2014.igem.org/Team:UMaryland/project/futureapplications" class="contentpagetitle"> <br />
Future Applications <br />
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<p>Future Applications of Research</p><br />
<p>We believe that the initial stages of the Biosensor have been complete and will lead to many new and exciting developments in the future. The general idea is that once the surface protein has been expressed, it will be able to interact with Dermo and produce a signal.</p><br />
<p>As for this year we our efforts working with a Bovine-Galectin have come the furthest and are ready to move into these next steps. Additionally in future years we hope to incorporate CvGal1 into the constructs, an effort we were not able to accomplish due to lack of time and troubleshooting. </p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong>Signaling Approaches:</strong></span></p><br />
<p> </p><br />
<p>1). Genetic engineering approach -Downstream Signaling Cascade</p><br />
<p><img src="https://static.igem.org/mediawiki/parts/9/99/UMSIgnal_Cascade.jpg" border="0" width="671" height="503" style="line-height: 12.0000009536743px; display: block; margin-left: auto; margin-right: auto; cursor: se-resize !important;" /></p><br />
<p>One of the strategies we hope to pursue is developing or incorporating a downstream signaling cascade into the cell strain. In this effort, once the E. coli came in contact with the Dermo it would then set off a transcription factor, ultimately producing a signal. Multiple approaches have been researched, including a Cpx membrane stress cascade, a split GFP cascade, and others.</p><br />
<p>The Cpx two component system reacts naturally to membrane stress. When the much larger Dermo is bound to the surface receptor expressed on E. coli, the bacterial cell membrane would become deformed. In turn, this would transmit a signal to the nucleus via the NlpE system that could be coupled to GFP production. This assay would require florescence microscopy to detect Dermo in solution.</p><br />
<p>Another option is for a split GFP construct. The idea is two different constructs containing the ligand are present in the cell, each with a different fraction of the GFP gene. When Dermo comes in close contact with the cell, multiple ligands on the cell would bind to the receptors on Dermo, causing the membrane to pinch or come together. This would trigger a spontaneous chance that two complementary pieces of GFP in close contact would bind, producing a fluorescent signal. This would also need to be observed using microscopy.</p><br />
<p>One of the ultimate goals from using the downstream signaling approach is to develop a “Kamikaze Bacteria”. By this we hope that the bacteria would respond to the presence of Dermo by eliminating it, possibly through killing itself. Similar work has previously been done with bees and colony collapse.</p><br />
<p>2). Microfluidics Approach- Size Based Cell sorter</p><br />
<p style="text-align: center;"><img src="https://static.igem.org/mediawiki/parts/0/02/UMMicrofluidics.png" border="0" width="572" height="416" /></p><br />
<p>Another avenue we plan to pursue is a microfluidics based detection approach. The overall concept would rely on the fact that E.coli cells are much smaller than Dermo cells. By creating a microfluidic device that sorts based on size, E.coli would travel into one output, whereas the Dermo and Dermo+E.coli complexes would travel into another output. Furthermore if the E.coli cells are fluorescently stained, analysis of the larger size fraction would indicate whether any E.coli cells are bound, confirming the presence of Dermo. While further differentiations based on size between bound and unbound Dermo may not be possible due to growth differences, this method will still suffice. </p><br />
<p>A microfluidic device would also contain the E.coli preventing them from entering the surrounding environment. Containment would be an obvious concern if a device like this were to ever hit the market. In this solution theoretically one could have pre-labeled E.coli in the machine stored and ready. The end user would simply intake a water sample right from the source, allow it to incubate, and process (sort) it. Collecting the larger fraction and viewing under a microscope would give a quick and easy real time method for detecting Dermo.</p><br />
<p>We believe that with the groundwork we have laid this year these future endeavors will be made possible as UMaryland continues to grow and gain expertise.</p><br />
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<option value="https://2014.igem.org/Team:UMaryland">&nbsp;Homepage</option><br />
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<option value="https://2014.igem.org/Team:UMaryland/meetourteam">&nbsp;Team</option><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/photos
Team:UMaryland/photos
2014-10-17T19:27:58Z
<p>Nbb826: </p>
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class="item116 level1"><span class="mymarg"><a class="yjanchor " href="https://igem.org/Team.cgi?id=1489"><span class="yjm_has_none"><span class="yjm_title">iGEM Page</span></span></a></span></li><li class="item117 level1"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/team/attributions"><span class="yjm_has_none"><span class="yjm_title">Attributions</span></span></a></span></li><li class="item300 level1 lilast"><span class="mymarg"><a class="yjanchor last" href="https://2014.igem.org/Team:UMaryland/team/sponsors"><span class="yjm_has_none"><span class="yjm_title">Sponsors</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li><li class="haschild item112 level0"><span class="child"><a class="yjanchor yjscroll" href="https://2014.igem.org/Team:UMaryland/project/overview"><span class="yjm_has_none"><span class="yjm_title">Project</span></span></a></span><ul class="subul_main level1"><li class="bl"></li><li class="tl"></li><li class="tr"></li><li class="item138 level1 first"><span class="mymarg"><a class="yjanchor first" href="https://2014.igem.org/Team:UMaryland/project/overview"><span class="yjm_has_none"><span class="yjm_title">Overview</span></span></a></span></li><li class="item145 level1"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/project/background"><span class="yjm_has_none"><span class="yjm_title">Background</span></span></a></span></li><li class="item144 level1"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/project/biobricks"><span class="yjm_has_none"><span class="yjm_title">parts (BioBricks)</span></span></a></span></li><li class="item102 level1"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/project/results" target="_blank" ><span class="yjm_has_none"><span class="yjm_title">Results</span></span></a></li><li class="item254 level1"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/project/futureapplications"><span class="yjm_has_none"><span class="yjm_title">future applications</span></span></a></span></li><li class="item301 level1 lilast"><span class="mymarg"><a class="yjanchor last" href="https://igem.org/Safety/Safety_Form?team_id=1489"><span class="yjm_has_none"><span class="yjm_title">Safety</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li><li class="haschild item234 level0"><span class="child"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/notebook"><span class="yjm_has_none"><span class="yjm_title">Notebook</span></span></a></span><ul class="subul_main level1"><li class="bl"></li><li class="tl"></li><li class="tr"></li><li class="item303 level1 first lilast"><span class="mymarg"><a class="yjanchor firstlast" href="https://2014.igem.org/Team:UMaryland/notebook"><span class="yjm_has_none"><span class="yjm_title">Wetlab Notebook</span></span></a></span></li><li 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<p><span style="line-height: 12.0000009536743px;">Chesapeake Bay Foundation Visit!</span></p><br />
<p><img src="https://static.igem.org/mediawiki/parts/a/a6/Bayfoundation1.jpg" border="0" width="300" height="234" style="border: 0;" /></p><br />
<p><span style="font-size: 11.0pt; line-height: 115%; font-family: 'Calibri','sans-serif'; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-hansi-theme-font: minor-latin; mso-bidi-font-family: 'Times New Roman'; mso-bidi-theme-font: minor-bidi; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;">Iowis Zhu and Sampada Koshatwar worked at the Chesapeake Bay Foundation’s Oyster Restoration/Oyster Gardening Center. </span></p><br />
<p><span style="font-family: Calibri, sans-serif; font-size: 14.4444446563721px; line-height: 16.8666667938232px;">Oyster Farm Visit</span></p><br />
<p><img src="https://static.igem.org/mediawiki/parts/8/8d/Oysterfarm.jpg" border="0" width="300" height="212" style="border: 0;" /></p><br />
<p> <span style="font-family: Calibri, sans-serif; font-size: 11pt; line-height: 115%;">From left: Iowis Zhu, Janine Taira, oyster farmer Kevin McClaren, Cami Uechi, and Adip Bhargav. The team spoke with an oyster farmer to learn more about the different perspectives surrounding the dermo infestation levels in the Chesapeake.</span></p><br />
<p> </p><br />
<p><span style="font-size: 11.0pt; line-height: 115%; font-family: 'Calibri','sans-serif'; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-hansi-theme-font: minor-latin; mso-bidi-font-family: 'Times New Roman'; mso-bidi-theme-font: minor-bidi; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;"> <img src="https://static.igem.org/mediawiki/parts/c/c8/Oysterfarm2.jpg" border="0" width="286" height="161" style="border: 0;" /></span><img src="https://static.igem.org/mediawiki/parts/5/53/Oysterfarm3.jpg" border="0" width="288" height="163" style="font-family: Calibri, sans-serif; font-size: 14.4444446563721px; line-height: 16.8666667938232px; cursor: se-resize !important; border: 0;" /></p><br />
<p><img src="https://static.igem.org/mediawiki/parts/6/65/Oysterfarm4.jpg" border="0" width="289" height="164" style="border: 0;" /></p><br />
<p>Churchill School Visit</p><br />
<p><img src="https://static.igem.org/mediawiki/parts/b/bb/Pavelschoolvisit.jpg" border="0" width="289" height="216" style="border: 0;" /></p><br />
<p><span style="font-size: 11.0pt; line-height: 115%; font-family: 'Calibri','sans-serif'; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-hansi-theme-font: minor-latin; mso-bidi-font-family: 'Times New Roman'; mso-bidi-theme-font: minor-bidi; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;">Junior Pavel Vlasov presented the project to students at Winston Churchill High School to foster interest in synthetic biology.</span></p><br />
<p><span style="font-size: 11.0pt; line-height: 115%; font-family: 'Calibri','sans-serif'; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-hansi-theme-font: minor-latin; mso-bidi-font-family: 'Times New Roman'; mso-bidi-theme-font: minor-bidi; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;"> </span></p><br />
<p>Collaboration Visits</p><br />
<p><img src="https://static.igem.org/mediawiki/parts/8/80/UmarylandVastavisit.jpg" border="0" width="291" height="163" /></p><br />
<p><span style="font-size: 11.0pt; line-height: 115%; font-family: 'Calibri','sans-serif'; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-hansi-theme-font: minor-latin; mso-bidi-font-family: 'Times New Roman'; mso-bidi-theme-font: minor-bidi; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;">From left: Dr. Gerardo Vasta, Janine Taira, Adip Bhargav, Iowis Zhu, and Andrew Zhao. The UMaryland iGem team met with Dr. Vasta, who discovered the CvGal1 receptor.</span></p><br />
<p><span style="font-size: 11.0pt; line-height: 115%; font-family: 'Calibri','sans-serif'; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-hansi-theme-font: minor-latin; mso-bidi-font-family: 'Times New Roman'; mso-bidi-theme-font: minor-bidi; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;"> </span></p><br />
<p>Visit to the DNR</p><br />
<p> <img src="https://static.igem.org/mediawiki/parts/a/a6/UMDNR1.jpg" border="0" width="226" height="169" /></p><br />
<p><img src="https://static.igem.org/mediawiki/parts/1/15/UMDNR2.jpg" border="0" width="219" height="292" /></p><br />
<p><img src="https://static.igem.org/mediawiki/parts/2/22/UMDNR3.jpg" border="0" width="219" height="164" /></p><br />
<p> </p><br />
<p><img src="https://static.igem.org/mediawiki/parts/9/99/UMDNR4.jpg" border="0" width="216" height="287" /></p><br />
<p><img src="https://static.igem.org/mediawiki/parts/c/c7/UMDNR5.jpg" border="0" width="214" height="286" /></p><br />
<p>Janine Taira, Aditya Biswas and Andrew Zhao all visited the Maryland Department of Natural Resources to learn more about Dermo, including current laboratory detection methods.</p> </div> <br />
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<option value="https://2014.igem.org/Team:UMaryland/meetourteam">&nbsp;Team</option><br />
<option value="https://2014.igem.org/Team:UMaryland/meetourteam">&nbsp;--Meet Our Team Members</option><br />
<option value="https://igem.org/Team.cgi?id=1489">&nbsp;--iGEM Page</option><br />
<option value="https://2014.igem.org/Team:UMaryland/team/attributions">&nbsp;--Attributions</option><br />
<option value="https://2014.igem.org/Team:UMaryland/team/sponsors">&nbsp;--Sponsors</option><br />
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<option value="https://2014.igem.org/Team:UMaryland/project/biobricks">&nbsp;--parts (BioBricks)</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/results">&nbsp;--Results</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/futureapplications">&nbsp;--future applications</option><br />
<option value="https://igem.org/Safety/Safety_Form?team_id=1489">&nbsp;--Safety</option><br />
<option value="#">&nbsp;Notebook</option><br />
<option value="https://2014.igem.org/Team:UMaryland/notebook">&nbsp;--Wetlab Notebook</option><br />
<option value="#">&nbsp;Human Practices</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/overview">&nbsp;--Overview | Mission Statement</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicschool">&nbsp;--Public School Outreach</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicschool">&nbsp;---HighSchool</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/community">&nbsp;--Community Interaction</option><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/project/results
Team:UMaryland/project/results
2014-10-17T19:25:09Z
<p>Nbb826: </p>
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<div>{{UMaryland/CSS}}<br />
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<h1 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/project/results" class="contentpagetitle"> <br />
Results <br />
</a><br />
</h1><br />
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<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/2014/e/e0/UMFig1-1.jpg" border="0" width="213px;" height="248px;" style="border: NaNpx solid black; transform: rotate(0.00rad); -webkit-transform: rotate(0.00rad);" /></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/2014/2/28/UMFig1-2.png" border="0" width="354px;" height="148px;" style="border: NaNpx solid black; transform: rotate(0.00rad); -webkit-transform: rotate(0.00rad);" /></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> A B</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig. 1 – A: Double RE digest (EcoRI and PstI) of pBAD-Bovine Galectin 1 and OmpA-Bovine Gibson assemblies. For pBAD-BtGal1, all lanes for after ladder have bands of the correct weight, while no lanes for OmpA-Bovine are of the correct weight. B: Expected band sizes after double digest for pBAD-Bovine Galectin 1. The EcoRI cut site in the Bovine Galectin 1 gene was subsequently removed via site directed mutagenesis.</span></p><br />
<p><strong id="docs-internal-guid-809a97bc-f600-9f66-fada-73accace1d98" style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Mammalian galectins typically have a simpler quaternary structure than invertebrate galectins; they are usually dimers while invertebrate galectins are usually tetramers. Bovine galectin 1, from Bos taurus, is a simpler galectin than CvGal1 and can thus be utilized as a model galectin to anchor in the E. coli outer membrane. In addition, while the crystal structure of CvGal1 and the exact binding ligand on P. marinus that CvGal1 binds to are still under investigation, the crystal structure and nature of bovine galectin 1 have been well studied. We have cloned Bovine galectin 1 into the pSB1C3 backbone via Gibson assembly. The bovine galectin 1 gene was ordered as a gBlock from IDT and PCR amplified for Gibson assembly. All Gibson products were verified via sequencing. </span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/2014/3/3f/UMFig2-1.jpg" border="0" width="233" height="331" style="border: 0;" />A<img src="https://static.igem.org/mediawiki/2014/e/ef/UMFig2-2.png" border="0" width="354px;" height="151px;" style="border: 0; transform: rotate(0.00rad); -webkit-transform: rotate(0.00rad);" /></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">B</span></p><br />
<p><strong style="font-weight: normal;"> (3rd Lane contains OmpA-Bovine product)</strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig. 2 - </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">OmpA-Bovine (cut with BamHI and HindIII, before SDM)</span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig. 2 – Double digest of OmpA-Bovine Galectin 1 (BamHI and HindIII) Lane 3 contains bands of the correct weight after digest B: Expected band sizes after double digest for pBAD-Bovine Galectin 1. Site directed mutagenesis was subsequently performed in order to remove an EcoRI site in the Bovine galectin 1 gene. Figure </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">2A</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> shows the Bovine galectin-1 assembled into pSB1C3-pBAD-RBS as a fusion protein with OmpA. When expressed in </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">E. coli</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">, the galectin should continue off the C-terminus of the OmpA-linker and thus be transported to the extracellular matrix since it is attached to the OmpA transport protein. After plasmid assembly transfer of plasmid into a DH5α </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">E. coli</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> cells strain, the plasmid was extracted and digested with EcoRI and PstI. The pattern of three bands appears again due to an additional EcoRI restriction site. </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 12.0000009536743px;"><span><img src="https://static.igem.org/mediawiki/2014/4/41/UMFig4-1.jpg" border="0" width="735" height="272" /></span></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 12.0000009536743px;">Sequencing Results:</p><br />
<p style="line-height: 12.0000009536743px;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$0:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$0:0">OmpA-Bovine SDM Chromatogram:</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$1:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$2:0">Sequence file:</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$3:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$4:0">NNNNNNNNNNNTNNNNANNNNNAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGGATG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$5:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$6:0">ATTTCTGGAATTCGCGGCCGCTTCTAGAGACATTGATTATTTGCACGGCGTCACACTTTGCTATGCCATAGCAAGATAGT</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$7:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$8:0">CCATAAGATTAGCGGATCCTACCTGACGCTTTTTATCGCAACTCTCTACTGTTTCTCCATACCGTTTTTTTGGGCTAGCT</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$9:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$10:0">ACTAGAGAAAGAGGAGAAATACTAGCATATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGC</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$11:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$12:0">TGGCAGGTTGCTCCAGCAACGCTAAAATCGATCAGGGAATTAACCCGTATGTTGGCTTTGAAATGGGTTACGACTGGTTA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$13:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$14:0">GGTCGTATGCCGTACAAAGGCAGCGTTGAAAACGGTGCATACAAAGCTCAGGGCGTTCAACTGACCGCTAAACTGGGTTA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$15:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$16:0">CCCAATCACTGACGACCTGGACATCTACACTCGTCTGGGTGGCATGGTATGGCGTGCAGACACTAAATCCAACGTTTATG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$17:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$18:0">GTAAAAACCACGACACCGGCGTTTCTCCGGTCTTCGCTGGCGGTGTTGAGTACGCGATCACTCCTGAAATCGCTACCCGT</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$19:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$20:0">CTGGAATACCAGTGGACCAACAACATCGGTGACGCACACACCATCGGCACTCGTCCGGACAACGGCGGANGTTCTGGAGG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$21:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$22:0">ANGGAGCATGGCGTCGGGGCTTGTTGCGTCCAATCTGAATCTGAAACCAGGTGAAAGCTTACGCGTCCGCGGCGAAGTAG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$23:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$24:0">CCGCAGATGCGAAATCTTTTCTCTTGAACCTGGGCAAGGACGANAATAATCTGGCTTTGCATTTCAACCCGCGTTTCNAC</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$25:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$26:0">GCTCATGGNGACNTGAACACAATCNTGTGTAANTCCAAGGNTGCTGGTGCTTGGGGTGCANAANNANCGNGNNNTCCGCG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$27:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$28:0">TTCCCGTTTCANNNNNGCTCANNNNGGNAGTCACCNTNCNGCNTTCANCCANNNCGANNTNCNNNNTTAAANNNGNNNNN</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$29:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$30:0">NGGNNNCNAAGTTNNANTTNNCNAANCNNNTNNAACNNNNNGNNNNNNNNNNNNNNNNNGNNNN</span></span></p><br />
<p style="line-height: 12.0000009536743px;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$0:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$30:0">A</span></span></p><br />
<p style="line-height: 12.0000009536743px;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$0:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$30:0"><img src="https://static.igem.org/mediawiki/2014/b/bc/UMFig5-1.jpg" border="0" width="733" height="272" /></span></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"> </p><br />
<p style="line-height: 12.0000009536743px;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$0:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$30:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$0:0">pBAD-Bovine SDM chromatogram:</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$1:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$2:0">NNNNNNNNNNNNTNNNCTATNNNNTAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$3:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$4:0">ATGATTTCTGGAATTCGCGGCCGCTTCTAGAGACATTGATTATTTGCACGGCGTCACACTTTGCTATGCCATAGCAAGAT</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$5:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$6:0">AGTCCATAAGATTAGCGGATCCTACCTGACGCTTTTTATCGCAACTCTCTACTGTTTCTCCATACCGTTTTTTTGGGCTA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$7:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$8:0">GCTACTAGAGAAAGAGGAGAAATACTAGATGGCGTCGGGGCTTGTTGCGTCCAATCTGAATCTGAAACCAGGTGAAAGCT</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$9:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$10:0">TACGCGTCCGCGGCGAAGTAGCCGCAGATGCGAAATCTTTTCTCTTGAACCTGGGCAAGGACGATAATAATCTGGCTTTG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$11:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$12:0">CATTTCAACCCGCGTTTCAACGCTCATGGCGACGTGAACACAATCGTGTGTAATTCCAAGGATGCTGGTGCTTGGGGTGC</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$13:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$14:0">AGAACAGCGTGAGTCCGCGTTCCCGTTTCAACCGGGCTCAGTTGTGGAAGTCACCATCAGCTTCAACCAGACCGATCTCA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$15:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$16:0">CCATTAAACTGCCGGATGGCTACGAGTTTAAGTTTCCAAACCGTTTAAACCTCGAGGCCATCAACTATCTGTCCGCGGGG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$17:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$18:0">GGCGACTTCAAGATTAAAAGCGTGCACCATCATCACCACCATTGATCACACTGGCTCACCTTCGGGTGGGCCTTTCTGCG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$19:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$20:0">TTTATATACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTTCTTTAAAACCGA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$21:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$22:0">AAAGATTACTTCGCGTTATGCNGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$23:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$24:0">GCTCACTCAAAGGCGGTAATACGGTTATCNNCAGAATCNGGGGATNACGCAGGAAANAACATGTGAGCAAANNNCAGCAA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$25:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$26:0">AANGCAGGAANCGTAAAANNNN</span></span></span></p><br />
<p style="line-height: 12.0000009536743px;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$0:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$30:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$26:0">B</span></span></span></p><br />
<p> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig. 3 - <strong>S</strong></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">equencing results and Chromatogram of SDM’d plasmids (</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">BBa_K1489000 and BBa_K1489004)</span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">To address this issue, site-directed mutagenesis was performed on these plasmids to change the EcoRI sequence within the bovine galectin-1 gene (5’-GAATTC-3’) to 5’-GAGTTT-3’, which will code for the same Glu-Phe sequence. Figure </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">3A</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> and </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">3B</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> demonstrate the two successfully mutagenized base pairs in the bovine galectin-1 gene, while preserving the rest of the plasmid.</span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p> <img src="https://static.igem.org/mediawiki/parts/a/a5/UMFinalfigureigem.jpg" border="0" width="687" height="269" style="line-height: 12.0000009536743px; cursor: se-resize !important;" /></p><br />
<p> Sequencing results:<br />
NNNNNNNNNNNNNNNNTATANAAATAGGCGTATCNCGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGG<br />
ATGATTTCTGGAATTCGCGGCCGCTTCTAGAGGGCATATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTT<br />
CTACTCTGCTGGCAGGTTGCTCCAGCAACGCTAAAATCGATCAGGGAATTAACCCGTATGTTGGCTTTGAAATGGGTTAC<br />
GACTGGTTAGGTCGTATGCCGTACAAAGGCAGCGTTGAAAACGGTGCATACAAAGCTCAGGGCGTTCAACTGACCGCTAA<br />
ACTGGGTTACCCAATCACTGACGACCTGGACATCTACACTCGTCTGGGTGGCATGGTATGGCGTGCAGACACTAAATCCA<br />
ACGTTTATGGTAAAAACCACGACACCGGCGTTTCTCCGGTCTTCGCTGGCGGTGTTGAGTACGCGATCACTCCTGAAATC<br />
GCTACCCGTCTGGAATACCAGTGGACCAACAACATCGGTGACGCACACACCATCGGCACTCGTCCGGACAACGGCGGAGG<br />
TTCTGGAGGAGGGAGCTCTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTT<br />
CTTTAAAACCGAAAAGATTACTTCGCGTTATGCAGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGG<br />
CGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCANGNNNAACATGTGAGCA<br />
AAAGGCCAGCNNAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCACAGGCTCCGCCCCCCTGACGAGCAT<br />
CACAAAAATCGACGCTCAAGTCAGANGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGNAGCTC<br />
CCTCGTGCGCTCTCCTGTTCCNACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTT<br />
CTCNTAGCTCACGCTGTAGGTATCNNCANNN</p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig 4. - <strong>S</strong></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">equencing Results and Chromatogram of OmpA moved into pSB1C3</span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Since UMaryland worked extensively with the OmpA-linker (BBa_K103006), we have also looked into improvements on that BioBrick.</span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; background-color: #f7f7f7;">OmpA-KasI chromatogram, shrunk down</span></span></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 12.0000009536743px;"><img src="https://static.igem.org/mediawiki/2014/0/0b/UMFig3-1.jpg" border="0" width="687" height="269" /></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 12.0000009536743px;">Sequencing Results:</p><br />
<p style="line-height: 12.0000009536743px;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$4:0">Sequence:</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$5:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$6:0">NNNNNNNNNNNNNNNTATAAAAATAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGGA</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$7:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$8:0">TGATTTCTGGAATTCGCGGCCGCTTCTAGAGGGCATATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTC</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$9:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$10:0">TACTCTGCTGGCAGGTTGCTCCAGCAACGCTAAAATCGATCAGGGAATTAACCCGTATGTTGGCTTTGAAATGGGTTACG</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$11:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$12:0">ACTGGTTAGGTCGTATGCCGTACAAAGGCAGCGTTGAAAACGGTGCATACAAAGCTCAGGGCGTTCAACTGACCGCTAAA</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$13:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$14:0">CTGGGTTACCCAATCACTGACGACCTGGACATCTACACTCGTCTGGGTGGCATGGTATGGCGTGCAGACACTAAATCCAA</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$15:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$16:0">CGTTTATGGTAAAAACCACGACACCGGCGTTTCTCCGGTCTTCGCTGGCGGTGTTGAGTACGCGATCACTCCTGAAATCG</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$17:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$18:0">CTACCCGTCTGGAATACCAGTGGACCAACAACATCGGTGACGCACACACCATCGGCACTCGTCCGGACAACGGCGGAGGT</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$19:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$20:0">TCTGGAGGAGGGGCGCCTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTTC</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$21:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$22:0">TTTAAAACCGAAAAGATTACTTCGCGTTATGCAGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGC</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$23:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$24:0">GAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCA</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$25:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$26:0">AAAGGNCAGCAAAAGGNCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCACAGGCTCCGCCCCCCTGACGAGCA</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$27:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$28:0">TCACAAAAATCGACGCTCAAGTCANNNNGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTC</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$29:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$30:0">CCTCGTGCGCTCTNCTGTTCCNACCCTGCCGCTTACCNGATACCTGTCNCNTTCTNCCTTCGGGANCGNGNGCTTTCTCA</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$31:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$32:0">TAGCTCACGCTNTAGGTANNNCANTNCGGNNNTNGNCGTTNNNCNCNANNCTNGNNTN</span></p><br />
<p> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig. 5 - <strong>S</strong></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">equencing Results and Chromatogram of SDM’d OmpA in pSB1C3</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><br /></span><span style="line-height: 12.777777671814px;">The previous version of Lpp-OmpA-Linker (BBa_K103006), designed for the pSB1A2 vector backbone, contained the non-traditional restriction sites NdeI (CATATG) and SacI (GAGCTC) in order to attach proteins at the N-terminus and C-terminus of OmpA, respectively. While the NdeI site is still useful for RE cloning at the N-terminus of OmpA, the implementation of the new plasmid backbone pSB1C3 has made the SacI site difficult to use. This is due to the presence of a second SacI site in the chloramphenicol acetyltransferase gene in pSB1C3. We have thus decided to mutagenize the previous SacI site on Lpp-OmpA-Linker into a KasI site in order to remove this issue. The KasI site (GGCGCC) encodes the amino acids glycine and alanine, converting the sequence of the linker from Gly-Gly-Gly-Ser-Gly-Gly-Gly-Ser to Gly-Gly-Gly-Ser-Gly-Gly-Gly-Ala. We believe this to be a sufficiently minor alteration to the unfolded linker sequence as to not affect protein function.</span></p> <br />
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<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; background-color: #f7f7f7;"></span></span></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 12.0000009536743px;"><img src="https://static.igem.org/mediawiki/parts/0/0c/UMBovinegalectinexpression.jpg" border="0" width="687" height="269" /></p><br />
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<p> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig. 6 - <strong>S</strong></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Western Blot of expression of SDM’d OmpA-BovineGalectin in pSB1C3</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><br /></span><span style="line-height: 12.777777671814px;">"Fig. 6 - Expression test for pBAD-Bovine galectin 1 and OmpA-Bovine galectin 1. Western Blot with anti-His antibody was done on soluble and insoluble fractions of non-induced cells and induced cells (0.2% arabinose for 21 hours). Bands can be observed in induced cells for pBAD-Bovine galectin 1 and both induced and non-induced cells for OmpA-Bovine galectin 1"</span></p><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/meetourteam/adip
Team:UMaryland/meetourteam/adip
2014-10-17T19:11:09Z
<p>Nbb826: </p>
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<h1>Meet our awesome team</h1><br />
<p> </p><br />
<p>Find our more about the three groups of contributors that made our project possible</p></div></div></div></div> </div><br />
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Adip Bhargav <br />
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<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/1/13/AdipBhargav.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate|Sophomore|Neurobiology &amp; Physiology</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
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<h4 class="yjtboxtitle">Adip Bhargav:</h4><br />
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<p> </p><br />
<span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px;">I am a junior majoring in Neurobiology and Physiology and minoring in French at the University of Maryland, College Park. At UMD, I am involved in many activities such as student government and teaching as an Undergraduate Teaching Assistant in Organic Chemistry. I plan on going to medical school after my undergraduate studies. I joined iGEM in order to experience all the different aspects of research. From fundraising to applying lab work to human practices, I've gained many invaluable skills through iGEM. When I'm not working on iGEM things, I love to meet new people and play tennis in my free time!.</span></div> <br />
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<div class="yjsg7_before"><div id="yjsg7" class="yjsg_grid yjsgsitew"><div id="user23" class="yjsgxhtml only_mod"><div class="yjsquare modid117"><div class="h2_holder"><h2 class="module_title"><span class="title_split titlesplit0">About</span> <span class="title_split titlesplit1">Umaryland</span></h2></div><div class="yjsquare_in"><p>UMaryland2014 is University of Maryland, College Parks, inaugural iGEM team. We are a combined effort of several departments and numerous faculty mentors. Although it is only our first year, believe our hard work and dedication has paid off. We can't wait for this years competition! GO TERPS!</p></div></div></div></div></div> <div class="footer_holders footer"><!-- footer --><br />
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<span class="yjmm_select" id="yjmm_selectid">Meet Our Team Members</span><br />
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<option value="/iGemWiki/">&nbsp;Homepage</option><br />
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<option value="https://2014.igem.org/Team:UMaryland/project/biobricks">&nbsp;--parts (BioBricks)</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/results">&nbsp;--Results</option><br />
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<option value="https://2014.igem.org/Team:UMaryland/notebook">&nbsp;--Wetlab Notebook</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/overview">&nbsp;Human Practices</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/overview">&nbsp;--Overview | Mission Statement</option><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/meetourteam
Team:UMaryland/meetourteam
2014-10-17T19:07:45Z
<p>Nbb826: </p>
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<h1>Meet our awesome team</h1><br />
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<p>Find our more about the three groups of contributors that made our project possible</p></div></div></div></div> </div><br />
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<a href="https://2014.igem.org/Team:UMaryland/meetourteam/tanya"><br />
Tanya Gordonov </a><br />
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<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/1/1d/TanyaGordonov.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
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<p style="text-align: center;"><strong>Graduate Advisor | Bioengineering</strong></p><br />
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<a href="https://2014.igem.org/Team:UMaryland/meetourteam/adip"><br />
Adip Bhargav </a><br />
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<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate | Junior | Neurobiology &amp; Physiology | French Minor</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
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<a href="https://2014.igem.org/Team:UMaryland/meetourteam/aditya"><br />
Aditya Biswas </a><br />
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<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/6/6d/Adityabiswas.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
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<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate | Junior | Bioengineering</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
<p> </p><br />
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<a href="https://2014.igem.org/Team:UMaryland/meetourteam/navadeep"><br />
Navadeep Boruah </a><br />
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<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/0/08/Navadeep2.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong>Graduate Advisor | Chemical and Biomolecular Engineering</strong></p><br />
<p> </p><br />
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<a href="https://2014.igem.org/Team:UMaryland/meetourteam/andrew"><br />
Andrew Zhao </a><br />
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<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/0/06/AndrewZhao.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
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<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate | Sophomore | Bioengineering</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
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<a href="https://2014.igem.org/Team:UMaryland/meetourteam/kevin"><br />
Kevin Pineault </a><br />
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<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/4/42/KevinPineault.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong>Undergraduate | Junior | Bioengineering</strong></p><br />
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<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/kevin"><br />
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<a href="https://2014.igem.org/Team:UMaryland/meetourteam/iowis"><br />
Iowis Zhu </a><br />
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<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/d/d1/IowisZhu.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
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<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate | Junior | Biochemistry/Cell Biology and Molecular Genetics</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
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<a href="https://2014.igem.org/Team:UMaryland/meetourteam/chris"><br />
Chris Euerle </a><br />
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<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/d/d6/Chriseuerle.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
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<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate | Junior | Computer Science and Neurobiology &amp; Physiology</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
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<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/chris"><br />
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<a href="https://2014.igem.org/Team:UMaryland/meetourteam/boots"><br />
Dr. Boots Quimby </a><br />
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<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/f/f0/Bootsquimby.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
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<p style="text-align: center;"><strong>Faculty Advisor | Integrated Life Sciences</strong></p><br />
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<a href="https://2014.igem.org/Team:UMaryland/meetourteam/janine"><br />
Janine Taira </a><br />
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<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/4/49/JanineTaira.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
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<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate | Sophomore | Neurobiology &amp; Physiology</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
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<a href="https://2014.igem.org/Team:UMaryland/meetourteam/camila"><br />
Camila Uechi </a><br />
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<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/6/63/Camilauechi.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
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<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate | Junior | Bioengineering</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
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<a href="https://2014.igem.org/Team:UMaryland/meetourteam/jason"><br />
Jason Kahn </a><br />
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<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/d/d1/JasonKahn.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
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<p style="text-align: center;"><strong>Faculty Advisor|Biochemistry</strong></p><br />
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<div class="first float-left width33"><br />
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<div class="news_item_c teamPage"><br />
<h2 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/meetourteam/nathan"><br />
Nathan Barber </a><br />
</h2><br />
<div class="clr"></div><br />
<div class="newsitem_text"><br />
<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/8/81/NathanBarber.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate | Senior | Bioengineering | Engineering Leadership Minor</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
<p> </p><br />
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<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/nathan"><br />
<span><br />
Read more: Nathan Barber </span><br />
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<a href="https://2014.igem.org/Team:UMaryland/meetourteam/pavel"><br />
Pavel Vlasov </a><br />
</h2><br />
<div class="clr"></div><br />
<div class="newsitem_text"><br />
<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/5/55/Pavelvlasov.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate | Sophomore | Biochemistry/Cell Biology &amp; Molecular Genetics</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
<p> </p><br />
</div><br />
<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/pavel"><br />
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Read more: Pavel Vlasov </span><br />
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<a href="https://2014.igem.org/Team:UMaryland/meetourteam/ed"><br />
Dr. Edward Eisenstein </a><br />
</h2><br />
<div class="clr"></div><br />
<div class="newsitem_text"><br />
<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/7/77/Edeisenstein.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong>Faculty Advisor | Bioengineering</strong></p><br />
<p> </p><br />
</div><br />
<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/ed"><br />
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Read more: Dr. Edward Eisenstein </span><br />
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<li class="first"><br />
<span class="catitem_title"><a href="https://2014.igem.org/Team:UMaryland/meetourteam/grad"><br />
Graduate Student Advisors</a><br />
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<span class="catitem_title"><a href="https://2014.igem.org/Team:UMaryland/meetourteam/undergrad"><br />
Undergraduate Students</a><br />
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</li><br />
<li class="last"><br />
<span class="catitem_title"><a href="https://2014.igem.org/Team:UMaryland/meetourteam/faculty"><br />
Faculty Advisors</a><br />
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<div class="yjsg7_before"><div id="yjsg7" class="yjsg_grid yjsgsitew"><div id="user23" class="yjsgxhtml only_mod"><div class="yjsquare modid117"><div class="h2_holder"><h2 class="module_title"><span class="title_split titlesplit0">About</span> <span class="title_split titlesplit1">Umaryland</span></h2></div><div class="yjsquare_in"><p>UMaryland2014 is University of Maryland, College Parks, inaugural iGEM team. We are a combined effort of several departments and numerous faculty mentors. Although it is only our first year, believe our hard work and dedication has paid off. We can't wait for this years competition! GO TERPS!</p></div></div></div></div></div> <div class="footer_holders footer"><!-- footer --><br />
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<span class="yjmm_select" id="yjmm_selectid">Meet Our Team Members</span><br />
<select id="mmenu" class="yjstyled"><br />
<option value="https://2014.igem.org/Team:UMaryland">&nbsp;Homepage</option><br />
<option value="https://2014.igem.org/Team:UMaryland/photos">&nbsp;Photos</option><br />
<option value="https://2014.igem.org/Team:UMaryland/meetourteam">&nbsp;Team</option><br />
<option value="https://2014.igem.org/Team:UMaryland/meetourteam" selected="selected">&nbsp;--Meet Our Team Members</option><br />
<option value="https://igem.org/Team.cgi?id=1489">&nbsp;--iGEM Page</option><br />
<option value="https://2014.igem.org/Team:UMaryland/team/attributions">&nbsp;--Attributions</option><br />
<option value="https://2014.igem.org/Team:UMaryland/team/sponsors">&nbsp;--Sponsors</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/overview">&nbsp;Project</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/overview">&nbsp;--Overview</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/background">&nbsp;--Background</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/biobricks">&nbsp;--parts (BioBricks)</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/results">&nbsp;--Results</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/futureapplications">&nbsp;--future applications</option><br />
<option value="https://igem.org/Safety/Safety_Form?team_id=1489">&nbsp;--Safety</option><br />
<option value="https://2014.igem.org/Team:UMaryland/notebook">&nbsp;Notebook</option><br />
<option value="https://2014.igem.org/Team:UMaryland/notebook">&nbsp;--Wetlab Notebook</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/overview">&nbsp;Human Practices</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/overview">&nbsp;--Overview | Mission Statement</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicschool">&nbsp;--Public School Outreach</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicschool">&nbsp;---HighSchool</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/community">&nbsp;--Community Interaction</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicawareness">&nbsp;--Public Awareness</option><br />
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Nbb826
http://2014.igem.org/Team:UMaryland
Team:UMaryland
2014-10-17T02:55:28Z
<p>Nbb826: </p>
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<li id="current" class=" active item101 level0 first"><span class="mymarg"><a class="yjanchor first activepath " href="https://2014.igem.org/Team:UMaryland"><span class="yjm_has_none"><span class="yjm_title">Homepage</span></span></a></span></li><li class="item104 level0"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/photos"><span class="yjm_has_none"><span class="yjm_title">Photos</span></span></a></span></li><li class="haschild item105 level0"><span class="child"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/meetourteam"><span class="yjm_has_none"><span class="yjm_title">Team</span></span></a></span><ul class="subul_main level1"><li class="bl"></li><li class="tl"></li><li class="tr"></li><li class="item293 level1 first"><span class="mymarg"><a class="yjanchor first" href="https://2014.igem.org/Team:UMaryland/meetourteam"><span class="yjm_has_none"><span class="yjm_title">Meet Our Team Members</span></span></a></span></li><li class="item116 level1"><span class="mymarg"><a class="yjanchor " href="https://igem.org/Team.cgi?id=1489"><span class="yjm_has_none"><span class="yjm_title">iGEM Page</span></span></a></span></li><li class="item117 level1"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/team/attributions"><span class="yjm_has_none"><span class="yjm_title">Attributions</span></span></a></span></li><li class="item300 level1 lilast"><span class="mymarg"><a class="yjanchor last" href="https://2014.igem.org/Team:UMaryland/team/sponsors"><span class="yjm_has_none"><span class="yjm_title">Sponsors</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li><li class="haschild item112 level0"><span class="child"><a class="yjanchor yjscroll" href="#"><span class="yjm_has_none"><span class="yjm_title">Project</span></span></a></span><ul class="subul_main level1"><li class="bl"></li><li class="tl"></li><li class="tr"></li><li class="item138 level1 first"><span class="mymarg"><a class="yjanchor first" href="https://2014.igem.org/Team:UMaryland/project/overview"><span class="yjm_has_none"><span class="yjm_title">Overview</span></span></a></span></li><li class="item145 level1"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/project/background"><span class="yjm_has_none"><span class="yjm_title">Background</span></span></a></span></li><li class="item144 level1"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/project/biobricks"><span class="yjm_has_none"><span class="yjm_title">parts (BioBricks)</span></span></a></span></li><li class="item102 level1"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/project/results" target="_blank" ><span class="yjm_has_none"><span class="yjm_title">Results</span></span></a></li><li class="item254 level1"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/project/futureapplications"><span class="yjm_has_none"><span class="yjm_title">future applications</span></span></a></span></li><li class="item301 level1 lilast"><span class="mymarg"><a class="yjanchor last" href="https://igem.org/Safety/Safety_Form?team_id=1489"><span class="yjm_has_none"><span class="yjm_title">Safety</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li><li class="haschild item234 level0"><span class="child"><a class="yjanchor " href="#"><span class="yjm_has_none"><span class="yjm_title">Notebook</span></span></a></span><ul class="subul_main level1"><li class="bl"></li><li class="tl"></li><li class="tr"></li><li class="item303 level1 first lilast"><span class="mymarg"><a class="yjanchor firstlast" href="https://2014.igem.org/Team:UMaryland/notebook"><span class="yjm_has_none"><span class="yjm_title">Wetlab Notebook</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li><li class="haschild item123 level0"><span class="child"><a class="yjanchor " href="#"><span class="yjm_has_none"><span class="yjm_title">Human Practices</span></span></a></span><ul class="subul_main level1"><li class="bl"></li><li class="tl"></li><li class="tr"></li><li class="item106 level1 first"><span class="mymarg"><a class="yjanchor first" href="https://2014.igem.org/Team:UMaryland/humanprac/overview"><span class="yjm_has_none"><span class="yjm_title">Overview | Mission Statement</span></span></a></span></li><li class="haschild item129 level1"><span class="child subparent"><a class="yjanchor grouptitle" href="https://2014.igem.org/Team:UMaryland/humanprac/publicschool"><span class="yjm_has_none"><span class="yjm_title">Public School Outreach</span></span></a></span><ul class="subul_main level2"><li class="bl"></li><li class="tl"></li><li class="tr"></li><li class="item134 level2 first lilast"><span class="mymarg"><a class="yjanchor firstlast" href="https://2014.igem.org/Team:UMaryland/humanprac/publicschool"><span class="yjm_has_none"><span class="yjm_title">HighSchool</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li><li class="item131 level1"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/humanprac/community"><span class="yjm_has_none"><span class="yjm_title">Community Interaction</span></span></a></span></li><li class="item132 level1 lilast"><span class="mymarg"><a class="yjanchor last" href="https://2014.igem.org/Team:UMaryland/humanprac/publicawareness"><span class="yjm_has_none"><span class="yjm_title">Public Awareness</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li></ul></div><br />
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<p>Home of The University of Maryland's first iGEM team.</p><br />
<p><a href="https://2014.igem.org/Team:UMaryland/project/overview"><br /><br /></a></p><br />
<p><a href="https://2014.igem.org/Team:UMaryland/project/overview" target="_self"><img src="https://static.igem.org/mediawiki/parts/5/55/UMIgem_logo_%281%29.png" border="0" alt="Our research heavily focuses on saving the Chesapeake Bay through protecting the oysters" width="1000" height="364" onmouseover="this.src='https://static.igem.org/mediawiki/parts/f/f2/Umarylandigeminfographic.jpg'" onmouseout="this.src= 'https://static.igem.org/mediawiki/parts/5/55/UMIgem_logo_%281%29.png'" style="border: 0;" /></a></p></div></div></div></div> </div><br />
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<h4>Project</h4><br />
<p><img src="http://static.guim.co.uk/sys-images/Lifeandhealth/Pix/pictures/2008/11/05/oyster1.jpg" border="0" width="80" height="48" /></p><br />
<a class="rm_content" href="https://2014.igem.org/Team:UMaryland/project/overview">Read More</a></div><br />
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<h4>Notebook</h4><br />
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<p><img src="https://static.igem.org/mediawiki/parts/9/97/UmarylandHumanpractices.png" border="0" width="99" height="72" style="border: 0;" /></p><br />
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<p><a href="https://2014.igem.org/Team:UMaryland/meetourteam" target="_self"><img src="https://static.igem.org/mediawiki/parts/7/7a/Umarylandlabphoto.jpg" border="0" alt="Our research heavily focuses on saving the Chesapeake Bay through protecting the oysters" width="500" height="150" onmouseover="this.src='https://static.igem.org/mediawiki/parts/d/d4/TeamPhoto.JPG'" onmouseout="this.src= 'https://static.igem.org/mediawiki/parts/7/7a/Umarylandlabphoto.jpg'" style="border: 0;" /></a></p></div></div></div></div></div> <div class="yjsg7_before"><div id="yjsg7" class="yjsg_grid yjsgsitew"><div id="user23" class="yjsgxhtml only_mod"><div class="yjsquare modid117"><div class="h2_holder"><h2 class="module_title"><span class="title_split titlesplit0">About</span> <span class="title_split titlesplit1">Umaryland</span></h2></div><div class="yjsquare_in"><p>UMaryland2014 is University of Maryland, College Parks, inaugural iGEM team. We are a combined effort of several departments and numerous faculty mentors. Although it is only our first year, believe our hard work and dedication has paid off. We can't wait for this years competition! GO TERPS!</p></div></div></div></div></div> <div class="footer_holders footer"><!-- footer --><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/humanprac/publicawareness
Team:UMaryland/humanprac/publicawareness
2014-10-16T22:32:11Z
<p>Nbb826: </p>
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<h1 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/humanprac/publicawareness" class="contentpagetitle"> <br />
Public Awareness <br />
</a><br />
</h1><br />
<br />
<br />
<br />
<p class="MsoNormal" style="text-align: center;"><strong><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"><span style="text-decoration: underline;">Public Awareness</span></span></strong></p><br />
<p> </p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">Realizing the importance of education and understanding, we strived to improve general understanding of <em>P. marinus</em> and the Dermo infection. We began at the University of Maryland with a survey to get students from all backgrounds to start thinking about the problems we were addressing. Next, we publicized our project and goals by connecting with deans of various colleges to spark interest in these fields. As awareness increased on campus, we turned our attention to the surrounding community. Networking with schools around the area, programs were established where iGEM members would go to the school and give a presentation or seminar tailored to the interests of the audience. For example, we gave a seminar on biotechnology and synthetic biology at a local high school in order to start education early with students who will be at the forefront of this type of research in the near future. The next step will be to get in touch with state organizations such as the Chesapeake Bay foundation in order to devise outreach initiatives and progressively raise awareness at higher levels of the community.</span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">Additionally we conducted a survey of over 250 students in undergraduate microbiology classes at the University of Maryland to further understand the public perception of this technology through the eyes of our next generation.The survey and responses are found below.</span></p><br />
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<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"> </span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">All responses to the survey will be kept anonymous, and no personal information about the participants will be released. The results of the survey may be published.</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;"> </span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">I have read and consent to the terms of confidentiality listed above ___________</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">1) What is a biosensor?</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">a. an organ that measures biological phenomenon</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">b. an organism that has the ability to sense other organisms</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">c. a device that measures biological parameters such as temperature or pressure</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><strong><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">d. an instrument that has a biological component and measures physical or chemical parameters</span></strong></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><strong><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;"> </span></strong></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">2) How do you feel about genetic engineering?</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">a. Strongly Support</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">b. Support</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">c. Neutral</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">d. Oppose</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">e. Strongly Oppose</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;"> </span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">3) How do you feel about the introduction of genetically modified </span><em style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">E. Coli</em><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;"> into the Chesapeake Bay?</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">a. Strongly Support</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">b. Support</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">c. Neutral</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">d. Oppose</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">e. Strongly Oppose</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;"> </span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">4) If it improved Chesapeake bay health and aquaculture?</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">a. Strongly Support</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">b. Support</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">c. Neutral</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">d. Oppose</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">e. Strongly Oppose</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;"> </span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">5) What danger do you think lies in using biotechnology and bioengineering to address environmental issues?</span></p><br />
<p class="MsoNormal"> </p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">(open ended)</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; text-align: center; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="text-decoration: underline;"><strong><span style="font-size: 10pt; font-family: Arial, sans-serif; color: #222222; text-decoration: underline;">ANSWER</span></strong></span><strong><span style="font-size: 10pt; font-family: Arial, sans-serif; color: #222222; text-decoration: underline;">S</span></strong></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;"><img src="https://static.igem.org/mediawiki/2014/1/15/UMSurveyAnswer1.jpg" border="0" width="365" height="268" style="float: left;" /></span><strong style="text-align: center;"><span style="font-size: 10pt; font-family: Arial, sans-serif; color: #222222; text-decoration: underline;"><img src="https://static.igem.org/mediawiki/2014/e/e9/UMSurveyAnswer2.jpg" border="0" width="410" height="268" style="cursor: se-resize !important; border: 0;" /></span></strong></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"> </p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><img src="https://static.igem.org/mediawiki/2014/0/07/UMSurveyAnswer3.jpg" border="0" width="365" height="268" style="border: 0;" /><img src="https://static.igem.org/mediawiki/2014/d/d2/UMSurveyAnswer4.jpg" border="0" width="365" height="268" /></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">By administering this survey to an undergraduate general microbiology class at the University of Maryland, we hoped to gain insight about where students stood on genetic engineering and its applications. We chose this class because it is a general education class at the University of Maryland which attracts students of various backgrounds and majors, not just science majors.</span></p><br />
<p class="MsoNormal"><span style="font-family: 'Times New Roman', serif; font-size: 12pt; line-height: 115%;"> With the first question, we tried to figure out just what proportion of the students were science majors or non-science majors, so that we could better interpret the responses to the later questions. We chose to ask what a biosensor was to determine if the student had significant background in biology or perhaps exposure to biotechnology. To our surprise, most students answered this question correctly which suggests that many students probably had some experience with or have heard about aspects of bioengineering. This conclusion reveals the extent of Science, Technology, Engineering, and Mathematics (STEM) education of University of Maryland students and the fact that most are fairly well informed about this type of technology.</span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">The next three questions corroborate the results from first question as shown by a relatively high number of students who are open to the idea of genetic engineering of <em>E. coli</em>. Even after being asked bluntly about the introduction of genetically-altered <em>E. coli</em> into the bay without any other information, more students, surprisingly, remained neutral rather than opposed. Finally, as expected, after qualifying the previous question by providing the motivation for the use of <em>E. coli</em>, a large shift was observed towards being supportive of the proposition. </span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"> Lastly, the free response questions (responses not shown) provided a deeper insight into the understanding of the population being surveyed. Most of the students developed valid points regarding possible areas of concern with this type of project. These ranged from ethical concerns to long-term, unexpected problems and the idea that native species may be adversely affected by our biosensor. It seemed to us that most of the concerns stemmed from a lack of understanding of the technology being used. Education and outreach efforts would likely eliminate some of these misunderstandings; however, some of the concerns raised are valid and are points we considered ourselves.</span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">The next step in this project would be to broaden the population being surveyed both in terms of size and diversity. In this way, we can determine if the results we obtained here is actually representative of a university setting. In terms of the participants being surveyed, we believe that young students are the most important because this is the generation that will be deciding on novel technologies that address pressing social and environmental concerns such as the health of the Chesapeake Bay. As a result, it is important to ensure that this demographic, especially, has access to correct information so that informed decisions can be made. However, we acknowledge that it is also important to gauge the views of current policy makers and people outside the university as well, so future surveys may be targeted at older demographics and different regions.</span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">With regards to the effect on public outreach efforts, it is clear that although most of the responses appeared to be informed there is room for improvement. Accordingly, we plan to continue our on campus and community efforts to advertise the issue and our research in order to start a thoughtful dialogue between those affected by concerns regarding the Chesapeake Bay and those who are trying to address these concerns.</span></p><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/humanprac/community
Team:UMaryland/humanprac/community
2014-10-16T22:31:27Z
<p>Nbb826: </p>
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class="item134 level2 first lilast"><span class="mymarg"><a class="yjanchor firstlast" href="https://2014.igem.org/Team:UMaryland/humanprac/publicschool"><span class="yjm_has_none"><span class="yjm_title">HighSchool</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li><li id="current" class=" active item131 level1"><span class="mymarg"><a class="yjanchor activepath " href="https://2014.igem.org/Team:UMaryland/humanprac/community"><span class="yjm_has_none"><span class="yjm_title">Community Interaction</span></span></a></span></li><li class="item132 level1 lilast"><span class="mymarg"><a class="yjanchor last" href="https://2014.igem.org/Team:UMaryland/humanprac/publicawareness"><span class="yjm_has_none"><span class="yjm_title">Public Awareness</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li></ul></div><br />
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Community Interaction <br />
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<p class="MsoNormal" style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">Interaction with Community</span></strong></span></p><br />
<p class="MsoNormal" style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"><img src="https://static.igem.org/mediawiki/parts/a/a5/Umarylandpanoramic.jpg" border="0" width="574" height="91" /> </span></strong></span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">We primarily focused our efforts on three organizations in the Chesapeake Bay community: Marinetics Choptank Oyster Farm, Department of Natural Resources, and Chesapeake Bay Foundation. Each provided insight into a different dimension of the Dermo infection problem. At Marinetics, we received an economy-centric view, where we learned a great deal about how <em>P. marinus</em> actually affects commercial oyster growth. We were surprised to find that, at least in that region of the bay, the owner of the farm experienced little to no effect from the pathogen. However, he did concede that detection systems for Dermo are not adequate and expressed that a real-time biosensor would be quite useful.</span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"> </span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"><img src="https://static.igem.org/mediawiki/parts/c/c8/Oysterfarm2.jpg" border="0" width="319" height="180" style="display: block; margin-left: auto; margin-right: auto;" /></span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">Next, we explored <em>P. marinus</em> through an environmental perspective as we spoke to members of the Chesapeake Bay Foundation. Here, we heard what was more expected of the problem including its adverse effects on oyster populations in the Chesapeake Bay. This interaction revealed the larger impacts our research would have on bay health as well as informed us about current approaches to remedy the problem. Still, we were unsure what to make of the two seemingly opposite views presented by the oyster farm and the Chesapeake Bay Foundation.</span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"> </span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"><img src="https://static.igem.org/mediawiki/parts/1/15/UMDNR2.jpg" border="0" width="180" height="241" style="display: block; margin-left: auto; margin-right: auto;" /></span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">Lastly, to clarify techniques and detection methods, we arranged a meeting with staff and lab members from the Department of Natural Resources. Mr. Chris Dungan, who is in charge of <em>P. marinus</em> investigation among other pathogens, gave us a comprehensive overview of how Dermo infections are detected and handled. We also discussed different aspects of policy-making and the obstacles to introducing new technology to combat these problems. For example, Mr. Dungan reconciled the apparently opposing accounts we received from the Chesapeake Bay Foundation and the oyster farm by telling us that Dermo infection is more prevalent in certain areas of the bay and will adversely affect populations in the bay but maybe not in aquaculture because oysters would be harvested and cooked before they exhibited the pathophysiology of the disease. Moreover, he acknowledged that oyster farmers tend to pay less attention to <em>P. marinus</em> in general because it is not a human pathogen. </span></p><br />
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<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">Ultimately, the fact that people tend to be apprehensive about new technologies and research that could be used to solve environmental problems motivated us to help address this issue. Since we understood just how powerful public opinion can be in promoting or labeling potential solutions prematurely as dangerous, we developed two main initiatives to promote education in these technologies and to gauge the public opinion quantitatively in our local community to understand why there is such reluctance towards new technology and the reasons behind this.</span></p> <br />
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<div class="yjsg7_before"><div id="yjsg7" class="yjsg_grid yjsgsitew"><div id="user23" class="yjsgxhtml only_mod"><div class="yjsquare modid117"><div class="h2_holder"><h2 class="module_title"><span class="title_split titlesplit0">About</span> <span class="title_split titlesplit1">Umaryland</span></h2></div><div class="yjsquare_in"><p>UMaryland2014 is University of Maryland, College Parks, inaugural iGEM team. We are a combined effort of several departments and numerous faculty mentors. Although it is only our first year, believe our hard work and dedication has paid off. We can't wait for this years competition! GO TERPS!</p></div></div></div></div></div> <div class="footer_holders footer"><!-- footer --><br />
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<option value="https://2014.igem.org/Team:UMaryland">&nbsp;Homepage</option><br />
<option value="https://2014.igem.org/Team:UMaryland/photos">&nbsp;Photos</option><br />
<option value="https://2014.igem.org/Team:UMaryland/meetourteam">&nbsp;Team</option><br />
<option value="https://2014.igem.org/Team:UMaryland/meetourteam">&nbsp;--Meet Our Team Members</option><br />
<option value="https://igem.org/Team.cgi?id=1489">&nbsp;--iGEM Page</option><br />
<option value="https://2014.igem.org/Team:UMaryland/team/attributions">&nbsp;--Attributions</option><br />
<option value="https://2014.igem.org/Team:UMaryland/team/sponsors">&nbsp;--Sponsors</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/overview">&nbsp;Project</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/overview">&nbsp;--Overview</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/background">&nbsp;--Background</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/biobricks">&nbsp;--parts (BioBricks)</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/results">&nbsp;--Results</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/futureapplications">&nbsp;--future applications</option><br />
<option value="https://igem.org/Safety/Safety_Form?team_id=1489">&nbsp;--Safety</option><br />
<option value="https://2014.igem.org/Team:UMaryland/notebook">&nbsp;Notebook</option><br />
<option value="https://2014.igem.org/Team:UMaryland/notebook">&nbsp;--Wetlab Notebook</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/overview">&nbsp;Human Practices</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/overview">&nbsp;--Overview | Mission Statement</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicschool">&nbsp;--Public School Outreach</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicschool">&nbsp;---HighSchool</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/community" selected="selected">&nbsp;--Community Interaction</option><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/humanprac/publicschool
Team:UMaryland/humanprac/publicschool
2014-10-16T22:29:23Z
<p>Nbb826: </p>
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School Outreach <br />
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<p> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-align: center;" dir="ltr"><span style="text-decoration: underline;"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: underline; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">Churchill High School</span></strong></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="text-decoration: underline;"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: underline; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;"> </span></strong></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="text-decoration: underline;"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: underline; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;"> </span></strong></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-align: center;" dir="ltr"><span style="text-decoration: underline;"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: underline; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;"><img src="https://static.igem.org/mediawiki/parts/b/bb/Pavelschoolvisit.jpg" border="0" width="494" height="371" style="border: 0;" /><br /></span></strong></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">In June of 2014, a member of our group conducted a short presentation about genetic engineering for an AP Biology class at Winston Churchill High School in Potomac, Maryland. The presentation contained information on the process of genetic engineering in general including transcription and translation, the basics of bacterial genetics, plasmid design; and on our team’s project including the importance of oysters to the Chesapeake Bay ecosystem and economy, the mechanism by which </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> infects oysters, and the protein structure being designed for our project. Approximately half of the class period, both during the presentation and after it, was left for students to ask questions. Some of the students were clearly interested in learning more about the current and future applications of genetic engineering, such as in agriculture, and about the possible risks involved and how they can be reduced. Some of the students asking these questions later said after the presentation that they were interested in studying genetics in college. Overall, the presentation was understandable and engaging to most of the class and several students were clearly interested in learning more about IGEM and our team’s current project.</span></p> <br />
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<div class="yjsg7_before"><div id="yjsg7" class="yjsg_grid yjsgsitew"><div id="user23" class="yjsgxhtml only_mod"><div class="yjsquare modid117"><div class="h2_holder"><h2 class="module_title"><span class="title_split titlesplit0">About</span> <span class="title_split titlesplit1">Umaryland</span></h2></div><div class="yjsquare_in"><p>UMaryland2014 is University of Maryland, College Parks, inaugural iGEM team. We are a combined effort of several departments and numerous faculty mentors. Although it is only our first year, believe our hard work and dedication has paid off. We can't wait for this years competition! GO TERPS!</p></div></div></div></div></div> <div class="footer_holders footer"><!-- footer --><br />
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<option value="https://2014.igem.org/Team:UMaryland">&nbsp;Homepage</option><br />
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<option value="https://2014.igem.org/Team:UMaryland/meetourteam">&nbsp;Team</option><br />
<option value="https://2014.igem.org/Team:UMaryland/meetourteam">&nbsp;--Meet Our Team Members</option><br />
<option value="https://igem.org/Team.cgi?id=1489">&nbsp;--iGEM Page</option><br />
<option value="https://2014.igem.org/Team:UMaryland/team/attributions">&nbsp;--Attributions</option><br />
<option value="https://2014.igem.org/Team:UMaryland/team/sponsors">&nbsp;--Sponsors</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/overview">&nbsp;Project</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/overview">&nbsp;--Overview</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/background">&nbsp;--Background</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/biobricks">&nbsp;--parts (BioBricks)</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/results">&nbsp;--Results</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/futureapplications">&nbsp;--future applications</option><br />
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<option value="https://2014.igem.org/Team:UMaryland/notebook">&nbsp;Notebook</option><br />
<option value="https://2014.igem.org/Team:UMaryland/notebook">&nbsp;--Wetlab Notebook</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/overview">&nbsp;Human Practices</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/overview">&nbsp;--Overview | Mission Statement</option><br />
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<option value="https://2014.igem.org/Team:UMaryland/humanprac/community">&nbsp;--Community Interaction</option><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/humanprac/overview
Team:UMaryland/humanprac/overview
2014-10-16T22:26:38Z
<p>Nbb826: </p>
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href="https://2014.igem.org/Team:UMaryland/project/futureapplications"><span class="yjm_has_none"><span class="yjm_title">future applications</span></span></a></span></li><li class="item301 level1 lilast"><span class="mymarg"><a class="yjanchor last" href="https://igem.org/Safety/Safety_Form?team_id=1489"><span class="yjm_has_none"><span class="yjm_title">Safety</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li><li class="haschild item234 level0"><span class="child"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/notebook"><span class="yjm_has_none"><span class="yjm_title">Notebook</span></span></a></span><ul class="subul_main level1"><li class="bl"></li><li class="tl"></li><li class="tr"></li><li class="item303 level1 first lilast"><span class="mymarg"><a class="yjanchor firstlast" href="https://2014.igem.org/Team:UMaryland/notebook"><span class="yjm_has_none"><span class="yjm_title">Wetlab Notebook</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li><li class=" active haschild item123 level0"><span class="child"><a class="yjanchor activepath " href="https://2014.igem.org/Team:UMaryland/humanprac/overview"><span class="yjm_has_none"><span class="yjm_title">Human Practices</span></span></a></span><ul class="subul_main level1"><li class="bl"></li><li class="tl"></li><li class="tr"></li><li id="current" class=" active item106 level1 first"><span class="mymarg"><a class="yjanchor first activepath " href="https://2014.igem.org/Team:UMaryland/humanprac/overview"><span class="yjm_has_none"><span class="yjm_title">Overview | Mission Statement</span></span></a></span></li><li class="haschild item129 level1"><span class="child subparent"><a class="yjanchor grouptitle" href="https://2014.igem.org/Team:UMaryland/humanprac/publicschool"><span class="yjm_has_none"><span class="yjm_title">Public School Outreach</span></span></a></span><ul class="subul_main level2"><li class="bl"></li><li 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Overviewmissionstatement <br />
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<p class="MsoNormal" style="text-align: center;"><strong><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"><span style="text-decoration: underline;">Overview Mission Statement</span></span></strong></p><br />
<p class="MsoNormal"><strong><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"> <img src="https://static.igem.org/mediawiki/parts/6/6c/Humanpracmain.png" border="0" width="593" height="553" style="display: block; margin-left: auto; margin-right: auto;" /></span></strong></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">From the beginning, our team wanted to pursue a project that would represent the spirit of Maryland. What better than to turn to the Chesapeake Bay, a natural resource that is priceless to Marylandians. The Chesapeake Bay, in addition to being a beautiful attraction, supports aquaculture and ecosystems that are essential to the economy and the environment. So, after choosing to address an oyster pathogen, one of many issues that threaten Chesapeake Bay health, we sought to understand the implications of our project and how it could be applied.</span></p><br />
<p> </p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">We organized various meetings with different Chesapeake Bay organizations to gauge the usefulness of our research and to further optimize our design. In doing so, we were made aware of the fact that public opinion played a large part in governing what research would be admissible and what the final product of the research could look like. Then, we took it to the next step to start our own initiatives to help educate and spread awareness about synthetic biology, and iGEM specifically in the context of the Chesapeake Bay and the environment. This outreach was accomplished through surveying a population of students and giving a class for middle school students.</span></p> <br />
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<div class="yjsg7_before"><div id="yjsg7" class="yjsg_grid yjsgsitew"><div id="user23" class="yjsgxhtml only_mod"><div class="yjsquare modid117"><div class="h2_holder"><h2 class="module_title"><span class="title_split titlesplit0">About</span> <span class="title_split titlesplit1">Umaryland</span></h2></div><div class="yjsquare_in"><p>UMaryland2014 is University of Maryland, College Parks, inaugural iGEM team. We are a combined effort of several departments and numerous faculty mentors. Although it is only our first year, believe our hard work and dedication has paid off. We can't wait for this years competition! GO TERPS!</p></div></div></div></div></div> <div class="footer_holders footer"><!-- footer --><br />
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<span class="yjmm_select" id="yjmm_selectid">Overview | Mission Statement</span><br />
<select id="mmenu" class="yjstyled"><br />
<option value="https://2014.igem.org/Team:UMaryland">&nbsp;Homepage</option><br />
<option value="https://2014.igem.org/Team:UMaryland/photos">&nbsp;Photos</option><br />
<option value="https://2014.igem.org/Team:UMaryland/meetourteam">&nbsp;Team</option><br />
<option value="https://2014.igem.org/Team:UMaryland/meetourteam">&nbsp;--Meet Our Team Members</option><br />
<option value="https://igem.org/Team.cgi?id=1489">&nbsp;--iGEM Page</option><br />
<option value="https://2014.igem.org/Team:UMaryland/team/attributions">&nbsp;--Attributions</option><br />
<option value="https://2014.igem.org/Team:UMaryland/team/sponsors">&nbsp;--Sponsors</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/overview">&nbsp;Project</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/overview">&nbsp;--Overview</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/background">&nbsp;--Background</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/biobricks">&nbsp;--parts (BioBricks)</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/results">&nbsp;--Results</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/futureapplications">&nbsp;--future applications</option><br />
<option value="https://igem.org/Safety/Safety_Form?team_id=1489">&nbsp;--Safety</option><br />
<option value="https://2014.igem.org/Team:UMaryland/notebook">&nbsp;Notebook</option><br />
<option value="https://2014.igem.org/Team:UMaryland/notebook">&nbsp;--Wetlab Notebook</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/overview">&nbsp;Human Practices</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/overview" selected="selected">&nbsp;--Overview | Mission Statement</option><br />
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<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicschool">&nbsp;---HighSchool</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/community">&nbsp;--Community Interaction</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicawareness">&nbsp;--Public Awareness</option><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/notebook
Team:UMaryland/notebook
2014-10-16T22:25:22Z
<p>Nbb826: </p>
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<h1 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/notebook" class="contentpagetitle"> <br />
Notebook <br />
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</h1><br />
<br />
<br />
<p class="MsoNormal"><strong>February to May </strong></p><br />
<p class="MsoNormal">Faculty and students interested in participating in UMaryland iGEM began meeting in February. Smaller interest meetings had occurred before to spread the word, as this is our first year! In these meetings, ideas were thrown around and brainstormed, and the logistics of participating in the iGEM competition were discussed. Every week for the first few months, a different student would give a presentation on a project and wiki from a previous year of iGEM in order to give all members a feel for what sort of projects have been done and what to expect throughout the process. In the later months, students presented then their own ideas for what UMaryland iGEM should pursue in 2014. Near the end of the semester, the group finally decided which of the proposed projects we would take – the development of a biosensor which would detect for the eastern oyster pathogen, <em>P. marinus</em>, in real-time using CvGal1, a galactoside-binding protein (galectin) found in the hemocytes of oysters, which has a strong affinity for binding to <em>P. marinus</em>.</p><br />
<p class="MsoNormal"> </p><br />
<p class="MsoNormal"><strong>May</strong></p><br />
<p class="MsoNormal">UMaryland iGEM began working in the lab at the end of May. Having chosen the outer membrane protein A (OmpA) to express the galectin on the surface of bacteria, the team ordered the part (BBa_K103006) and transformed it into DHα <em>E. coli</em> to create a stock of this plasmid. Team members also learned and refreshed on standard lab practices and protocols which would carry throughout most of the project: chemical transformation and bacterial culture, aseptic technique, minipreparation of a plasmid, and gel electrophoresis. From gel electrophoresis of the miniprep samples, cut at the unique BioBrick prefix and suffix with restriction enzymes EcoRI and PstI, as well as sequencing done by GENEWIZ, Inc., we could confirm that the OmpA plasmid was successfully cloned. This procedure would become our standard for cloning and confirming the identity of all subsequent plasmids.</p><br />
<p class="MsoNormal"> </p><br />
<p class="MsoNormal"><strong>June</strong></p><br />
<p class="MsoNormal">The team also began mapping out the BioBrick constructs intended for this year.</p><br />
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<p class="MsoNormal">The two constructs presented above differ only by the promoter sequence: the pBAD promoter is activated by arabinose, while the pLAC promoter is activated by IPTG. For this reason, the team also prepared a stock of pSB1C3-pBAD-RBS-GFP (BBa_K750000) and pSB1C3-pLAC-RBS-RFP (BBa_J04450) plasmids. By removing the fluorescent protein genes and linearizing the DNA, we would use these vectors as the basis for all the clones we intended to express, as the promoter, ribosomal binding site, and terminator are already part of the BioBrick pSB1C3 backbone.</p><br />
<p class="MsoNormal">Appropriately-designed primers were ordered from Integrated DNA Technologies (IDT) for preparation of Gibson assembly. With the primers, we PCR amplified the OmpA gene from its plasmid and the two vectors mentioned previously. Gels confirmed the unique sizes of the PCR products, and a NanoDrop spectrophotometer analyzed the DNA concentrations. The use of a NanoDrop machine would also become a standard in our DNA analysis, particularly for Gibson assembly, where concentrations are vital.</p><br />
<p class="MsoNormal">Due to issues with acquiring the CvGal1 gene, the team began a side project in the meantime. β-galactosidase, readily available as a BioBrick, was considered in place of CvGal1. Instead of binding a galactose sugar, as the galectin would, β-galactosidase would instead cleave it into monosaccharides. The LacZα (BBa_K329005) and full-length LacZ (BBa_K1155007) genes were prepared similar to OmpA, with the end result being PCR products of the isolated, desired LacZ gene. Since we were working with mainly DH5α <em>E. coli</em>, which have the LacZΩ fragment already present, the LacZα fragment expressed in DH5α is expected to combine and express the whole β-galactosidase. This mechanism was the preferred method, as the LacZα fragment is only around 240 bp in size. The full-length LacZ gene (approx. 3 kbp) was also used, however, in the event that the two-fragment method did not work.</p><br />
<p class="MsoNormal">The team also moved the OmpA gene from its original backone, the old pSB1A2 (ampicillin resistance), to the current BioBrick standard, pSB1C3 (chloramphenicol resistance). Using standard restriction enzyme cloning techniques and the pSB1C3 linearized backbone supplied by iGEM, the team was able to “cut-and-paste” the OmpA gene into its new vector. The process was fairly simple, due to the different antibiotic resistances acting as a general screen between the old and new plasmids. This was our first BioBrick made (BBa_K1489002), classifying as an improvement to an existing part.</p><br />
<p class="MsoNormal"> </p><br />
<p class="MsoNormal"><strong>July</strong></p><br />
<p class="MsoNormal">Going into July, the team continued this LacZ side project. We encountered many roadblocks when attempting to clone the LacZ gene into the pSB1C3-pBAD-RBS vector (pBAD was eventually chosen as the promoter of choice for the rest of our project) via Gibson reactions. The only viable plasmid produced, screened through blue colonies on X-gal-coated plate, was an attempt at cloning OmpA-LacZα. However, both the gel and sequencing results revealed a serious error within the OmpA gene.</p><br />
<p class="MsoNormal">Around the same time, at the end of July, we had received both CvGal1 and bovine galectin-1 gene fragments, ordered from IDT’s gBlock system. Both gBlocks were based on the wildtype genes, but modified to ensure a proper codon reading frame and to remove potential problems with the expressed protein, such as disulfide bonds forming due to the oxidative nature of the extracellular matrix. Immediately, the team PCR amplified all the fragments for Gibson assembly, where we faced an unresponsive CvGal1 gene. Every other fragment looked correct on the gel, however, so the team proceeded with focusing solely on bovine galectin-1 while troubleshooting CvGal1.</p><br />
<p class="MsoNormal"> </p><br />
<p class="MsoNormal"><strong>August</strong></p><br />
<p class="MsoNormal">August was filled with many trials, but two main goals: 1) Gibson cloning of the bovine galectin gene into the pSB1C3-pBAD-RBS backbone, one with fusion to the OmpA gene and one without, and 2) PCR amplification of the CvGal1 gBlock.</p><br />
<p class="MsoNormal">For the first goal, the team faced many issues with residual plasmid transforming, making it unclear whether the Gibson reaction was assembling the gene fragments properly. Finally, on August 15, it seemed we had at last cloned the bovine galectin into the vector (without OmpA). (Though done in parallel, the OmpA-Bovine galectin construct did not clone properly.) The gel revealed the plasmid’s identity clearly, but for the wrong reason – the bovine galectin gene had an EcoRI site in the middle of its sequence which had gone unnoticed, until this product was cloned and analyzed.</p><br />
<p class="MsoNormal">As for the CvGal1 gBlock, after various PCR attempts in which the team tested different Mg<sup>2+</sup> concentrations, DMSO concentrations, cycle time and temperature conditions, and DNA polymerases, the team contacted IDT about the inability to amplify the gene fragment. To the time of this writing, they are still in the process of remaking the CvGal1 gBlock, as the persistent issue appeared to be on their end.</p><br />
<p class="MsoNormal"> </p><br />
<p class="MsoNormal"><strong>September</strong></p><br />
<p class="MsoNormal">With the school semester starting again, our team members had much less time to spend in the lab. However, we continued to make progress, and toward the end of September, we had successfully cloned the OmpA-Bovine galectin construct as well. Persistence, fine-tuned procedures, and a bit of luck paid dividends as the sequencing results came back as a clear match.</p><br />
<p class="MsoNormal">The case of the unwanted EcoRI site remained within the bovine galectin genes, but the team was ready with a site-directed mutagenesis kit and primers. Mutagenesis on the EcoRI site to create a silent mutation was successful on its first try, much to our relief, and a second round of sequencing revealed that the EcoRI site in the bovine galectin gene in both constructs had been removed (but the BioBrick prefix’s EcoRI site remained intact).</p><br />
<p> </p><br />
<p class="MsoNormal">These are our two main project BioBricks: the Bovine galectin alone in pSB1C3-pBAD-RBS (BBa_K1489000) and the Bovine galectin gene fused to the end of the OmpA gene in the same vector (BBa_K1489004). Additionally, since the pSB1C3 backbone has a SacI site (which pSB1A2 did not) and OmpA has a SacI site (as designed by the creators of that BioBrick), the pSB1C3-OmpA which we created back in June had two unusable SacI sites. In order to maintain the integrity of that BioBrick with its unique restriction sites (along with the standard EcoRI and PstI sites), we also performed SDM on the pSB1C3-OmpA (BBa_K1489002) to replace the now-useless SacI site at the end of the OmpA gene with a KasI site, creating our fourth BioBrick (BBa_K1489003).</p><br />
<p class="MsoNormal"><strong>October</strong></p><br />
<p class="MsoNormal">With the BioBricks made, the team began preliminary protein expression tests. Multiple cultures of DH5α E. coli, transformed with either the bovine galectin or OmpA-BvGal1 BioBricks, were grown up to a noticeable OD. They were then induced with arabinose at increasing concentrations, from no induction to 0.2% m/v. After further incubation of the bacteria to allow for protein production, a 1 mL sample of each culture was pelleted. The cells were then lysed by resuspension in 20 μL SDS and boiling the solution, yielding a soluble protein fraction in the supernatant after centrifugation. The remaining pellet was boiled in 100 μL SDS with harsher conditions in order to extract as much insoluble protein as possible. These protein samples (soluble and insoluble fractions) were run through SDS-PAGE, followed by western blotting (taking advantage of the His6 tag placed in the synthesized bovine galectin-1). Anti-mouse was used for the secondary antibody. Expression of the bovine galectin BioBrick (BBa_K1489000) appeared strong and at the correct size, but expression of OmpA-BvGal1 (BBa_K1489004) featured weak bands in both the induced and uninduced samples, though of correct size.</p> <br />
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Future Applications <br />
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<p>Future Applications of Research</p><br />
<p>We believe that the initial stages of the Biosensor have been complete and will lead to many new and exciting developments in the future. The general idea is that once the surface protein has been expressed, it will be able to interact with Dermo and produce a signal.</p><br />
<p>As for this year we our efforts working with a Bovine-Galectin have come the furthest and are ready to move into these next steps. Additionally in future years we hope to incorporate CvGal1 into the constructs, an effort we were not able to accomplish due to lack of time and troubleshooting. </p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong>Signaling Approaches:</strong></span></p><br />
<p> </p><br />
<p>1). Genetic engineering approach -Downstream Signaling Cascade</p><br />
<p><img src="https://static.igem.org/mediawiki/parts/9/99/UMSIgnal_Cascade.jpg" border="0" width="671" height="503" style="line-height: 12.0000009536743px; display: block; margin-left: auto; margin-right: auto; cursor: se-resize !important;" /></p><br />
<p>One of the strategies we hope to pursue is developing or incorporating a downstream signaling cascade into the cell strain. In this effort, once the E. coli came in contact with the Dermo it would then set off a transcription factor, ultimately producing a signal. Multiple approaches have been researched, including a Cpx membrane stress cascade, a split GFP cascade, and others.</p><br />
<p>The Cpx two component system reacts naturally to membrane stress. When the much larger Dermo is bound to the surface receptor expressed on E. coli, the bacterial cell membrane would become deformed. In turn, this would transmit a signal to the nucleus via the Cpx system that could be coupled to GFP production. This assay would require florescence microscopy to detect Dermo in solution.</p><br />
<p>Another option is for a split GFP construct. The idea is two different constructs containing the ligand are present in the cell, each with a different fraction of the GFP gene. When Dermo comes in close contact with the cell, multiple ligands on the cell would bind to the receptors on Dermo, causing the membrane to pinch or come together. This would trigger a spontaneous chance that two complementary pieces of GFP in close contact would bind, producing a fluorescent signal. This would also need to be observed using microscopy.</p><br />
<p>One of the ultimate goals from using the downstream signaling approach is to develop a “Kamikaze Bacteria”. By this we hope that the bacteria would respond to the presence of Dermo by eliminating it, possibly through killing itself. Similar work has previously been done with bees and colony collapse.</p><br />
<p>2). Microfluidics Approach- Size Based Cell sorter</p><br />
<p style="text-align: center;"><img src="https://static.igem.org/mediawiki/parts/0/02/UMMicrofluidics.png" border="0" width="572" height="416" /></p><br />
<p>Another avenue we plan to pursue is a microfluidics based detection approach. The overall concept would rely on the fact that E.coli cells are much smaller than Dermo cells. By creating a microfluidic device that sorts based on size, E.coli would travel into one output, whereas the Dermo and Dermo+E.coli complexes would travel into another output. Furthermore if the E.coli cells are fluorescently stained, analysis of the larger size fraction would indicate whether any E.coli cells are bound, confirming the presence of Dermo. While further differentiations based on size between bound and unbound Dermo may not be possible due to growth differences, this method will still suffice. </p><br />
<p>A microfluidic device would also contain the E.coli preventing them from entering the surrounding environment. Containment would be an obvious concern if a device like this were to ever hit the market. In this solution theoretically one could have pre-labeled E.coli in the machine stored and ready. The end user would simply intake a water sample right from the source, allow it to incubate, and process (sort) it. Collecting the larger fraction and viewing under a microscope would give a quick and easy real time method for detecting Dermo.</p><br />
<p>We believe that with the groundwork we have laid this year these future endeavors will be made possible as UMaryland continues to grow and gain expertise.</p><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/project/results
Team:UMaryland/project/results
2014-10-16T22:22:54Z
<p>Nbb826: </p>
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<a href="https://2014.igem.org/Team:UMaryland/project/results" class="contentpagetitle"> <br />
Results <br />
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</h1><br />
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<br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/2014/e/e0/UMFig1-1.jpg" border="0" width="213px;" height="248px;" style="border: NaNpx solid black; transform: rotate(0.00rad); -webkit-transform: rotate(0.00rad);" /></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/2014/2/28/UMFig1-2.png" border="0" width="354px;" height="148px;" style="border: NaNpx solid black; transform: rotate(0.00rad); -webkit-transform: rotate(0.00rad);" /></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> A B</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig. 1 - Bovine Galectin-1 gene cloned into the pSB1C3 vector with pBAD promoter and a RBS</span></p><br />
<p><strong id="docs-internal-guid-809a97bc-f600-9f66-fada-73accace1d98" style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Galectin-1 from </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Bos taurus</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> has been cloned into pSB1C3 vector. </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Bos taurus </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">or bovine galectin enabled us to generate a platform that enables future work on ligand binding. While CvGals binding characteristics are still being researched the crystal structure and nature of bovine galectin has been well studied. The subsequent BioBricks regarding this gene were all constructed via Gibson assembly and cloning. The plasmids from which the OmpA insert and the vector were amplified via extension PCR are respectively: OmpA with linker (BBa_K103006) and pBAD-RBS on the pSB1C3 backbone (BBa_K750000). For the latter, the GFP in the BioBrick was not included for Gibson assembly. The bovine galectin-1 was ordered as a synthetically synthesized sequence from IDT as a linear gene and amplified directly with appropriate primers for Gibson assembly. </span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Figure </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">1A</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> shows the Bovine galectin-1 assembled into pSB1C3-pBAD-RBS alone (no OmpA), which when expressed in </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">E. coli</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> should produce a soluble form of the protein. After successful assembly and cloned into DH5α </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">E. coli</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">. The plasmid was cut with restriction enzymes EcoRI and PstI, standard sites in the pSB1C3 vector’s BioBrick prefix and suffix, respectively. The result is expected to yield the insert, pBAD-RBS-Bovine galectin-1, and the pSB1C3 vector. However, an unwanted EcoRI site was found in the galectin-1 gene afterwards, yielding the pattern seen in the gel. Figure </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">1B</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> demonstrates the expected pattern, with band sizes of 2033 bp, 496 bp, and 169 bp top to bottom, via the software “A plasmid Editor” (ApE, courtesy of M. Wayne Davis, University of Utah).</span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/2014/3/3f/UMFig2-1.jpg" border="0" width="233" height="331" style="border: 0;" />A<img src="https://static.igem.org/mediawiki/2014/e/ef/UMFig2-2.png" border="0" width="354px;" height="151px;" style="border: 0; transform: rotate(0.00rad); -webkit-transform: rotate(0.00rad);" /></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">B</span></p><br />
<p><strong style="font-weight: normal;"> (3rd Lane contains OmpA-Bovine product)</strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig. 2 - </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">OmpA-Bovine (cut with BamHI and HindIII, before SDM)</span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Figure </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">2A</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> shows the Bovine galectin-1 assembled into pSB1C3-pBAD-RBS as a fusion protein with OmpA. When expressed in </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">E. coli</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">, the galectin should continue off the C-terminus of the OmpA-linker and thus be transported to the extracellular matrix since it is attached to the OmpA transport protein. After plasmid assembly transfer of plasmid into a DH5α </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">E. coli</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> cells strain, the plasmid was extracted and digested with EcoRI and PstI. The pattern of three bands appears again due to an additional EcoRI restriction site. </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 12.0000009536743px;"><span><img src="https://static.igem.org/mediawiki/2014/4/41/UMFig4-1.jpg" border="0" width="735" height="272" /></span></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 12.0000009536743px;">Sequencing Results:</p><br />
<p style="line-height: 12.0000009536743px;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$0:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$0:0">OmpA-Bovine SDM Chromatogram:</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$1:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$2:0">Sequence file:</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$3:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$4:0">NNNNNNNNNNNTNNNNANNNNNAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGGATG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$5:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$6:0">ATTTCTGGAATTCGCGGCCGCTTCTAGAGACATTGATTATTTGCACGGCGTCACACTTTGCTATGCCATAGCAAGATAGT</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$7:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$8:0">CCATAAGATTAGCGGATCCTACCTGACGCTTTTTATCGCAACTCTCTACTGTTTCTCCATACCGTTTTTTTGGGCTAGCT</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$9:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$10:0">ACTAGAGAAAGAGGAGAAATACTAGCATATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGC</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$11:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$12:0">TGGCAGGTTGCTCCAGCAACGCTAAAATCGATCAGGGAATTAACCCGTATGTTGGCTTTGAAATGGGTTACGACTGGTTA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$13:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$14:0">GGTCGTATGCCGTACAAAGGCAGCGTTGAAAACGGTGCATACAAAGCTCAGGGCGTTCAACTGACCGCTAAACTGGGTTA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$15:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$16:0">CCCAATCACTGACGACCTGGACATCTACACTCGTCTGGGTGGCATGGTATGGCGTGCAGACACTAAATCCAACGTTTATG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$17:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$18:0">GTAAAAACCACGACACCGGCGTTTCTCCGGTCTTCGCTGGCGGTGTTGAGTACGCGATCACTCCTGAAATCGCTACCCGT</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$19:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$20:0">CTGGAATACCAGTGGACCAACAACATCGGTGACGCACACACCATCGGCACTCGTCCGGACAACGGCGGANGTTCTGGAGG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$21:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$22:0">ANGGAGCATGGCGTCGGGGCTTGTTGCGTCCAATCTGAATCTGAAACCAGGTGAAAGCTTACGCGTCCGCGGCGAAGTAG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$23:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$24:0">CCGCAGATGCGAAATCTTTTCTCTTGAACCTGGGCAAGGACGANAATAATCTGGCTTTGCATTTCAACCCGCGTTTCNAC</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$25:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$26:0">GCTCATGGNGACNTGAACACAATCNTGTGTAANTCCAAGGNTGCTGGTGCTTGGGGTGCANAANNANCGNGNNNTCCGCG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$27:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$28:0">TTCCCGTTTCANNNNNGCTCANNNNGGNAGTCACCNTNCNGCNTTCANCCANNNCGANNTNCNNNNTTAAANNNGNNNNN</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$29:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$30:0">NGGNNNCNAAGTTNNANTTNNCNAANCNNNTNNAACNNNNNGNNNNNNNNNNNNNNNNNGNNNN</span></span></p><br />
<p style="line-height: 12.0000009536743px;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$0:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$30:0">A</span></span></p><br />
<p style="line-height: 12.0000009536743px;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$0:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$30:0"><img src="https://static.igem.org/mediawiki/2014/b/bc/UMFig5-1.jpg" border="0" width="733" height="272" /></span></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"> </p><br />
<p style="line-height: 12.0000009536743px;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$0:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$30:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$0:0">pBAD-Bovine SDM chromatogram:</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$1:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$2:0">NNNNNNNNNNNNTNNNCTATNNNNTAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$3:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$4:0">ATGATTTCTGGAATTCGCGGCCGCTTCTAGAGACATTGATTATTTGCACGGCGTCACACTTTGCTATGCCATAGCAAGAT</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$5:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$6:0">AGTCCATAAGATTAGCGGATCCTACCTGACGCTTTTTATCGCAACTCTCTACTGTTTCTCCATACCGTTTTTTTGGGCTA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$7:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$8:0">GCTACTAGAGAAAGAGGAGAAATACTAGATGGCGTCGGGGCTTGTTGCGTCCAATCTGAATCTGAAACCAGGTGAAAGCT</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$9:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$10:0">TACGCGTCCGCGGCGAAGTAGCCGCAGATGCGAAATCTTTTCTCTTGAACCTGGGCAAGGACGATAATAATCTGGCTTTG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$11:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$12:0">CATTTCAACCCGCGTTTCAACGCTCATGGCGACGTGAACACAATCGTGTGTAATTCCAAGGATGCTGGTGCTTGGGGTGC</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$13:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$14:0">AGAACAGCGTGAGTCCGCGTTCCCGTTTCAACCGGGCTCAGTTGTGGAAGTCACCATCAGCTTCAACCAGACCGATCTCA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$15:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$16:0">CCATTAAACTGCCGGATGGCTACGAGTTTAAGTTTCCAAACCGTTTAAACCTCGAGGCCATCAACTATCTGTCCGCGGGG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$17:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$18:0">GGCGACTTCAAGATTAAAAGCGTGCACCATCATCACCACCATTGATCACACTGGCTCACCTTCGGGTGGGCCTTTCTGCG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$19:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$20:0">TTTATATACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTTCTTTAAAACCGA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$21:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$22:0">AAAGATTACTTCGCGTTATGCNGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$23:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$24:0">GCTCACTCAAAGGCGGTAATACGGTTATCNNCAGAATCNGGGGATNACGCAGGAAANAACATGTGAGCAAANNNCAGCAA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$25:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$26:0">AANGCAGGAANCGTAAAANNNN</span></span></span></p><br />
<p style="line-height: 12.0000009536743px;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$0:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$30:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$26:0">B</span></span></span></p><br />
<p> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig. 3 - <strong>S</strong></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">equencing results and Chromatogram of SDM’d plasmids (</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">BBa_K1489000 and BBa_K1489004)</span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">To address this issue, site-directed mutagenesis was performed on these plasmids to change the EcoRI sequence within the bovine galectin-1 gene (5’-GAATTC-3’) to 5’-GAGTTT-3’, which will code for the same Glu-Phe sequence. Figure </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">3A</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> and </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">3B</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> demonstrate the two successfully mutagenized base pairs in the bovine galectin-1 gene, while preserving the rest of the plasmid.</span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p> <img src="https://static.igem.org/mediawiki/parts/a/a5/UMFinalfigureigem.jpg" border="0" width="687" height="269" style="line-height: 12.0000009536743px; cursor: se-resize !important;" /></p><br />
<p> Sequencing results:<br />
NNNNNNNNNNNNNNNNTATANAAATAGGCGTATCNCGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGG<br />
ATGATTTCTGGAATTCGCGGCCGCTTCTAGAGGGCATATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTT<br />
CTACTCTGCTGGCAGGTTGCTCCAGCAACGCTAAAATCGATCAGGGAATTAACCCGTATGTTGGCTTTGAAATGGGTTAC<br />
GACTGGTTAGGTCGTATGCCGTACAAAGGCAGCGTTGAAAACGGTGCATACAAAGCTCAGGGCGTTCAACTGACCGCTAA<br />
ACTGGGTTACCCAATCACTGACGACCTGGACATCTACACTCGTCTGGGTGGCATGGTATGGCGTGCAGACACTAAATCCA<br />
ACGTTTATGGTAAAAACCACGACACCGGCGTTTCTCCGGTCTTCGCTGGCGGTGTTGAGTACGCGATCACTCCTGAAATC<br />
GCTACCCGTCTGGAATACCAGTGGACCAACAACATCGGTGACGCACACACCATCGGCACTCGTCCGGACAACGGCGGAGG<br />
TTCTGGAGGAGGGAGCTCTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTT<br />
CTTTAAAACCGAAAAGATTACTTCGCGTTATGCAGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGG<br />
CGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCANGNNNAACATGTGAGCA<br />
AAAGGCCAGCNNAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCACAGGCTCCGCCCCCCTGACGAGCAT<br />
CACAAAAATCGACGCTCAAGTCAGANGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGNAGCTC<br />
CCTCGTGCGCTCTCCTGTTCCNACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTT<br />
CTCNTAGCTCACGCTGTAGGTATCNNCANNN</p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig 4. - <strong>S</strong></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">equencing Results and Chromatogram of OmpA moved into pSB1C3</span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Since UMaryland worked extensively with the OmpA-linker (BBa_K103006), we have also looked into improvements on that BioBrick.</span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; background-color: #f7f7f7;">OmpA-KasI chromatogram, shrunk down</span></span></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 12.0000009536743px;"><img src="https://static.igem.org/mediawiki/2014/0/0b/UMFig3-1.jpg" border="0" width="687" height="269" /></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 12.0000009536743px;">Sequencing Results:</p><br />
<p style="line-height: 12.0000009536743px;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$4:0">Sequence:</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$5:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$6:0">NNNNNNNNNNNNNNNTATAAAAATAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGGA</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$7:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$8:0">TGATTTCTGGAATTCGCGGCCGCTTCTAGAGGGCATATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTC</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$9:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$10:0">TACTCTGCTGGCAGGTTGCTCCAGCAACGCTAAAATCGATCAGGGAATTAACCCGTATGTTGGCTTTGAAATGGGTTACG</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$11:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$12:0">ACTGGTTAGGTCGTATGCCGTACAAAGGCAGCGTTGAAAACGGTGCATACAAAGCTCAGGGCGTTCAACTGACCGCTAAA</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$13:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$14:0">CTGGGTTACCCAATCACTGACGACCTGGACATCTACACTCGTCTGGGTGGCATGGTATGGCGTGCAGACACTAAATCCAA</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$15:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$16:0">CGTTTATGGTAAAAACCACGACACCGGCGTTTCTCCGGTCTTCGCTGGCGGTGTTGAGTACGCGATCACTCCTGAAATCG</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$17:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$18:0">CTACCCGTCTGGAATACCAGTGGACCAACAACATCGGTGACGCACACACCATCGGCACTCGTCCGGACAACGGCGGAGGT</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$19:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$20:0">TCTGGAGGAGGGGCGCCTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTTC</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$21:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$22:0">TTTAAAACCGAAAAGATTACTTCGCGTTATGCAGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGC</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$23:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$24:0">GAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCA</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$25:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$26:0">AAAGGNCAGCAAAAGGNCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCACAGGCTCCGCCCCCCTGACGAGCA</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$27:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$28:0">TCACAAAAATCGACGCTCAAGTCANNNNGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTC</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$29:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$30:0">CCTCGTGCGCTCTNCTGTTCCNACCCTGCCGCTTACCNGATACCTGTCNCNTTCTNCCTTCGGGANCGNGNGCTTTCTCA</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$31:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$32:0">TAGCTCACGCTNTAGGTANNNCANTNCGGNNNTNGNCGTTNNNCNCNANNCTNGNNTN</span></p><br />
<p> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig. 5 - <strong>S</strong></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">equencing Results and Chromatogram of SDM’d OmpA in pSB1C3</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><br /></span><span style="line-height: 12.777777671814px;">The previous version of Lpp-OmpA-Linker (BBa_K103006), designed for the pSB1A2 vector backbone, contained the non-traditional restriction sites NdeI (CATATG) and SacI (GAGCTC) in order to attach proteins at the N-terminus and C-terminus of OmpA, respectively. While the NdeI site is still useful for RE cloning at the N-terminus of OmpA, the implementation of the new plasmid backbone pSB1C3 has made the SacI site difficult to use. This is due to the presence of a second SacI site in the chloramphenicol acetyltransferase gene in pSB1C3. We have thus decided to mutagenize the previous SacI site on Lpp-OmpA-Linker into a KasI site in order to remove this issue. The KasI site (GGCGCC) encodes the amino acids glycine and alanine, converting the sequence of the linker from Gly-Gly-Gly-Ser-Gly-Gly-Gly-Ser to Gly-Gly-Gly-Ser-Gly-Gly-Gly-Ala. We believe this to be a sufficiently minor alteration to the unfolded linker sequence as to not affect protein function.</span></p> <br />
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<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; background-color: #f7f7f7;"></span></span></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 12.0000009536743px;"><img src="https://static.igem.org/mediawiki/parts/0/0c/UMBovinegalectinexpression.jpg" border="0" width="687" height="269" /></p><br />
<br />
<p> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig. 6 - <strong>S</strong></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Western Blot of expression of SDM’d OmpA-BovineGalectin in pSB1C3</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><br /></span><span style="line-height: 12.777777671814px;">"Both the bovine galectin and the OmpA-Bovine Galectin were tested for expression using a Western blot. Each sample was also tested with and without induction. Additionally for all trials both the soluable and insoluable fractions were examined. The results showed expression of both the Bovine galectin and the OmpA-Bovine galectin. The conclusion points to sucessful expression of our biobrick "</span></p><br />
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<option value="https://2014.igem.org/Team:UMaryland/humanprac/overview">&nbsp;--Overview | Mission Statement</option><br />
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<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicschool">&nbsp;---HighSchool</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/community">&nbsp;--Community Interaction</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicawareness">&nbsp;--Public Awareness</option><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/project/background
Team:UMaryland/project/background
2014-10-16T22:21:37Z
<p>Nbb826: </p>
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<h1 class="article_title"><br />
<a href="/https://2014.igem.org/Team:UMaryland/project/background" class="contentpagetitle"> <br />
Background-Motivation <br />
</a><br />
</h1><br />
<br />
<br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: center;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"><em><span style="text-decoration: underline;"><strong>Background</strong></span></em><br /></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">We at UMaryland iGEM are developing a biosensor to detect for the presence of an oyster pathogen, </span><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">Perkinsus marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">. This parasite is commonly found over the eastern shore of North America, including in Maryland’s own Chesapeake Bay. </span><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">, a unicellular protist, causes an oyster infection known as Dermo (Espinosa, Winnicki, &amp; Allam, 2013). The pathogen has been shown to infest oysters by entering their hemocytes via induced phagocytosis. When inside the hemocyte, the parasite is capable of resisting the reactive oxygen species generated by the oyster immune system and modulating the apoptosis of infected hemocyte cells to spread itself further (Espinosa et al., 2013). This leads to proliferation inside the oyster hemocyte, eventually lysing the host cell and releasing more </span><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"> (Allam et al., 2013). This process leads to massive tissue damage and, ultimately, the death of the oyster.</span></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">It is believed that </span><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">P. marinus </span><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">enters utilizing a galectin, a galactose-binding protein, on the hemocyte cell surface, CvGal1, which allows it to bind preferentially to </span><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"> (Schott, Pecher, Okafor, &amp; Vasta, 2003). Our aim is to exploit this property by expressing the galectin on the surface of </span><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">E. coli</span><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"> bacterial cells by attaching it to the C-terminus of an OmpA transmembrane protein. These bacteria will thus bind to any </span><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"> present in a sample of water. Our long-term goal is to incorporate these biosensors into a lab-on-a-chip device, requiring only small samples and allowing quick response times for detecting this pathogen.</span></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">This year, we have focused on working with the bovine galectin-1 in place of CvGal1. At only 14 kDa in size, the bovine galectin was chosen as a simpler candidate for expression on the surface of the membrane (Liao, Kapadia, Ahmed, Vasta, &amp; Herzberg, 1994). Furthermore, this bovine galectin-1 was one of three galectins used for the homology modeling of CvGal1 due to their similar binding domains (Tasumi &amp; Vasta, 2007). Though bovine galectin-1 is a two-domain galectin, compared to the four domains of CvGal1 (Feng et al., 2013), it is believed that the bovine galectin has potential to bind the same ligand present on </span><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"> that CvGal1 binds. Additionally, we have optimized the bovine galectin-1 gene, most notably for better protein folding and BioBrick friendliness.</span></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">Although laboratory methods for Dermo detection using PCR exist, they are currently not capable of detecting the disease in real time. In the time needed to conduct a laboratory test, Dermo will have easily spread to new locations. If we can pinpoint the location of Dermo on-site, then we can take action ultimately to eliminate the parasite from eastern oyster habitats.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">References</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">Allam, B., Carden, W. E., Ward, J. E., Ralph, G., Winnicki, S., &amp; Espinosa, E. P. (2013). Early host-pathogen interactions in marine bivalves: Evidence that the alveolate parasite Perkinsus marinus infects through the oyster mantle during rejection of pseudofeces. </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">Journal of Invertebrate Pathology</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">, </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">113</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">(1), 26-34.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">Espinosa, E. P., Winnicki, S., &amp; Allam, B. (2013). Early host–pathogen interactions in a marine bivalve: Crassostrea virginica pallial mucus modulates Perkinsus marinus growth and virulence. </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">Diseases of Aquatic Organisms</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">, </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">104</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">(3), 237-247.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">Feng, C., Ghosh, A., Amin, M. N., Giomarelli, B., Shridhar, S., Banerjee, A., ... &amp; Vasta, G. R. (2013). The galectin CvGal1 from the eastern oyster (Crassostrea virginica) binds to blood group A oligosaccharides on the hemocyte surface. </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">Journal of Biological Chemistry</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">, </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">288</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">(34), 24394-24409.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">Liao, D. I., Kapadia, G., Ahmed, H., Vasta, G. R., &amp; Herzberg, O. (1994). Structure of S-lectin, a developmentally regulated vertebrate beta-galactoside-binding protein. </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">Proceedings of the National Academy of Sciences</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">, </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">91</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">(4), 1428-1432.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">Tasumi, S., &amp; Vasta, G. R. (2007). A galectin of unique domain organization from hemocytes of the Eastern oyster (Crassostrea virginica) is a receptor for the protistan parasite Perkinsus marinus. </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">The Journal of Immunology</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">, </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">179</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">(5), 3086-3098.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">Schott, E. J., Pecher, W. T., Okafor, F., &amp; Vasta, G. R. (2003). The protistan parasite Perkinsus marinus is resistant to selected reactive oxygen species. </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">Experimental Parasitology</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">, </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">105</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">(3-4), 232-240.</span></p><br />
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Nbb826
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Team:UMaryland/project/overview
2014-10-16T22:19:48Z
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Overview <br />
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<p style="margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: center;" dir="ltr"><em><span style="text-decoration: underline;"><strong><span style="vertical-align: baseline;"><span style="color: #000000; font-family: Arial;"><span style="font-size: 15.5555562973022px; line-height: 19.1666679382324px; white-space: pre-wrap;">Overview</span></span></span></strong></span></em></p><br />
<p style="margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: center;" dir="ltr"><em><span style="text-decoration: underline;"><strong><span style="vertical-align: baseline;"><span style="color: #000000; font-family: Arial;"><span style="font-size: 15.5555562973022px; line-height: 19.1666679382324px; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/parts/f/f2/Umarylandigeminfographic.jpg" border="0" width="402" height="256" /></span></span></span></strong></span></em></p><br />
<p style="margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: left;" dir="ltr"><span style="vertical-align: baseline;"><span style="color: #000000; font-family: Arial;"><span style="font-size: 15.5555562973022px; line-height: 19.1666679382324px; white-space: pre-wrap;">The UMaryland iGem team opted for a project that hits close to home; with the Chesapeake Bay in our backyard, we have the potential to make a direct impact on the community. As Maryland’s most important natural landmark, the Chesapeake Bay is an integral part of our state’s identity. The bay is home to a diverse ecosystem, and is an economic boon to the area, supporting both the seafood industry and local tourism. Since the health of the bay is important, we looked into current problems in need of attention. In particular, the predicament of the Chesapeake’s oysters piqued our interest. As oyster populations declined over the years, the absence of this filtration system left a host of problems, including decreased water clarity and rising chemical pollutant levels. If the oyster populations can be maintained, it will improve the overall health of the Chesapeake. One way to combat oyster fatalities is to develop a way to monitor Dermo infestations. </span></span> </span></p><br />
<p style="text-align: left;"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
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<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: center;" dir="ltr"><em><strong><span style="text-decoration: underline;"><span style="font-size: 15px; font-family: Arial; color: #000000; font-variant: normal; text-decoration: underline; vertical-align: baseline; white-space: pre-wrap; background-color: #ffffff;">Applications in the lab</span></span></strong></em></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">It is believed that </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">P. marinus </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">enters oyster hemocytes utilizing a galectin on the cell’s surface, CvGal1, that allows binds preferentially to </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">. Our aim is to exploit CvGal1’s ability to bind to </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> in order to design a biosensor for the parasite. By expressing the galectin on the surface of </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">E. coli</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> and triggering a signal transduction cascade within the bacterial cell, the </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">E. coli</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> could produce Green Fluorescent Protein. Thus, </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">P. marinus </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">binding to CvGal1 would trigger the expression of GFP within </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">E. coli, </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> creating a visual marker for the parasite. The majority of this project will be accomplished through genetic cloning and bacterial transformation. Although laboratory methods for Dermo detection using Polymerase Chain Reaction exist, they are currently not capable of detecting the disease in real time. In the time needed to conduct a laboratory test, Dermo could have easily spread to new locations. This biosensor would help to accurately pinpoint Dermo infestation levels in real time. If we can pinpoint the location of Dermo, as elucidated in this research proposal, then we can ultimately develop a biosensor that can react to Dermo with targeted destruction. </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"><strong id="docs-internal-guid-809a97bc-f5c8-6033-4262-354a17c98c1d" style="font-weight: normal;"><br /><br /></strong></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: center;" dir="ltr"><span style="text-decoration: underline;"><strong><em><span style="font-size: 15px; font-family: Arial; color: #000000; font-variant: normal; text-decoration: underline; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">Brief overview of servicing the community</span></em></strong></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">We having been traveling around Maryland conducting various community outreach efforts by visiting local schools and oyster farms. During each visit, members present our project and spread awareness on the current state of the Chesapeake Bay, as well as ways individuals can help service the Bay. Some presentations are made on synthetic biology to increase student participation and interest in the sciences. We also have a committee dedicated to creating an educational computer game that teaches others about the issues that the Chesapeake Bay currently faces. It is our team’s hope that our research results to be presented at the iGEM jamboree will have a profound and positive impact on the health of the Chesapeake Bay and oyster populations worldwide</span></p><br />
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<option value="https://2014.igem.org/Team:UMaryland">&nbsp;Homepage</option><br />
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<option value="https://2014.igem.org/Team:UMaryland/meetourteam">&nbsp;Team</option><br />
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<option value="https://2014.igem.org/Team:UMaryland/humanprac/overview">&nbsp;Human Practices</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/overview">&nbsp;--Overview | Mission Statement</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicschool">&nbsp;--Public School Outreach</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicschool">&nbsp;---HighSchool</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/community">&nbsp;--Community Interaction</option><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/team/attributions
Team:UMaryland/team/attributions
2014-10-16T22:16:01Z
<p>Nbb826: </p>
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<h1 style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">We Would like to thank all who have helped make this year possible!</span></h1><br />
<h2 style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> <br />A Special Thanks to the following contributors: </span></h2><br />
<p><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/parts/0/0e/Bentley.jpg" border="0" width="155" height="232" style="border: 0;" /></span></p><br />
<h3 style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">Dr. William Bentley</span></strong></h3><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="color: #000000; font-family: Arial; font-size: 15px; font-weight: normal; line-height: 1.15; text-indent: 36pt; white-space: pre-wrap; background-color: transparent;"> Distinguished professor and chair, Fischell Department of Bioengineering University of Maryland</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="color: #000000; font-family: Arial; font-size: 15px; font-weight: normal; line-height: 1.15; text-indent: 36pt; white-space: pre-wrap; background-color: transparent;">Dr. Bentley was helpful talking through the initial stages of the project, representing the schools Bioengineering department and getting out name out to campus, as well as a major supporter of the project, in addition to providing lab instruments, resources, and offering cellular constructs.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="color: #000000; font-family: Arial; font-size: 15px; font-weight: normal; line-height: 1.15; text-indent: 36pt; white-space: pre-wrap; background-color: transparent;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="color: #000000; font-family: Arial; font-size: 15px; font-weight: normal; line-height: 1.15; text-indent: 36pt; white-space: pre-wrap; background-color: transparent;"><img src="https://static.igem.org/mediawiki/parts/a/a6/JohnBuchner.jpg" border="0" width="126" height="188" style="border: 0;" /></span></p><br />
<h3 style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">Dr. John Buchner</span></strong></h3><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Lecturer, Department of Cell Biology and Molecular Genetics University of Maryland.</span><span style="line-height: 12.0000009536743px;">John Buchner is lecturer and lab coordinator for general microbiology, and a lab coordinator for microbial genetics. His microbiology class is in I Series courses at UMD. He is a member of the HPI group, a faculty collaboration intent improving the educational experience of undergraduates, and publishing the findings. John is also a Chesapeake Fellow, and is dedicated raising awareness of sustainability to the students he teaches, and reducing material consumed in his classes.</span></p><br />
<p class="MsoNormal"> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Dr. Buchner was instrumental in helping our students design the project, providing many insights into how proteins function through the cell membranes, as well as signal transduction pathways.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/parts/4/48/BenWoodard.jpg" border="0" width="145" height="239" /></span></p><br />
<h3 style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">Dr. Ben Woodard</span></strong></h3><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Director, Biotechnology Research and Education Program</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Director, Bioprocess Scale-up Facility</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Dr. Woodard was a valuable research throughout the planning phase of the project, providing insight on the potential commercial aspect of the product and keeping our team on track with realistic expectations.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<h3 style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">John Wilhelm III</span></strong></h3><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="color: #000000; font-family: Arial; font-size: 15px; line-height: 17.25px; white-space: pre-wrap;">Regional Sales manager at Integrated DNA technologies</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="color: #000000; font-family: Arial; font-size: 15px; line-height: 17.25px; white-space: pre-wrap;">Mr. Wilhelm provided a wealth of knowledge regarding the creation of our constructs and the different options availible, as well as introducing us to synthetic gene construction. IDT was responsible for creating our G-Blocks used for CVGAL1 and Bovine Galectin-1</span></p><br />
<p><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span><img src="https://static.igem.org/mediawiki/parts/8/8a/Vastaheadshot.jpg" border="0" alt="Gerardo R Vasta " width="175" height="219" style="border: 0;" /></p><br />
<h3 style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><strong>Dr. Gerardo Vasta</strong></span></h3><br />
<p><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Professor, Department of Microbiology and Immunology University of Maryland School of Medicine</span></p><br />
<p><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Dr. Vasta was extremely helpful with our questions regarding Dermo, specifically the ligand CVGAL1 and the interaction between Dermo and E.Coli as well providing strategies for expressing and optimizing this protein.</span></p><br />
<p><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/parts/1/15/Herzberg.jpg" border="0" width="180" height="190" style="border: 0;" /></span></p><br />
<h3 style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><strong>Dr. Osnat Herzberg</strong></span></h3><br />
<p><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Professor Department of Chemistry and Biochemistry University of Maryland at Shady Grove</span></p><br />
<p><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Dr. Herzberg contributed to our protein design, offering to check our sequences and provided insights into the bovine-galectin that we created.</span></p><br />
<h2><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">A Special Thanks to the Following Departments/Laboratories:</span></h2><br />
<h3 style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">The Cell Biology and Molecular Genetics</span></strong></h3><br />
<p><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">A special thanks for Cell Biology and Molecular Genetics for offering us a lab space throughout the duratation of the summer in which we conducted our research.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<h3 style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">Dr. Kahn's Laboratory</span></strong></h3><br />
<p><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;"><br /></span></strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">A special thanks to The Kahn Lab group for offering us lab space throughout the fall semester, providing additional materials we needed for research and offering storage space. Additionally Dr. Kahn and Dr. Quimby performed the ran the final western blot which was included in our results</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">Dr. Bentley's Laboratory</span></strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong style="line-height: 1.15;"><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;"><br /></span></strong><span style="color: #000000; font-family: Arial;"><span style="font-size: 15px; line-height: 17.25px; white-space: pre-wrap;">A special thanks to The Bentley Lab group for permitting us the use of their facilities,</span></span><span style="color: #000000; font-family: Arial; font-size: 15px; line-height: 17.25px; white-space: pre-wrap;">including multiple analysis machines without which we could not have completed our research,</span><span style="font-size: 15px; line-height: 17.25px; white-space: pre-wrap; color: #000000; font-family: Arial; text-indent: 36pt;"> providing enzymes and other research materials.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; line-height: 17.25px; white-space: pre-wrap; color: #000000; font-family: Arial; text-indent: 36pt;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">Dr. Eisensteins Laboratory</span></strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;"><br /></span></strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">A special thanks to the Eisenstein Lab group for providing us with research materials, as well as allowing us to run experiments at their location</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;"> </span></p><br />
<h3 style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">Fischell Department of Bioengineering</span></strong></h3><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">A. James Clark School of Engineering</span></strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">College of Computer, Math, and Natural Sciences</span></strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;"><span style="line-height: 1.15; text-indent: 36pt; background-color: transparent;">Integrated Life Sciences (ILS)</span></span></strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"> </p><br />
<h3><strong><span id="docs-internal-guid-809a97bc-c77f-0ff5-d846-9d15e5944245"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;"> National Scholarships Office (Leadership and Discovery Grant)</span></span></strong></h3> </div><br />
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<option value="https://2014.igem.org/Team:UMaryland">&nbsp;Homepage</option><br />
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<option value="https://2014.igem.org/Team:UMaryland/meetourteam">&nbsp;--Meet Our Team Members</option><br />
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<option value="https://2014.igem.org/Team:UMaryland/project/biobricks">&nbsp;--parts (BioBricks)</option><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/meetourteam
Team:UMaryland/meetourteam
2014-10-16T22:14:42Z
<p>Nbb826: </p>
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href="https://2014.igem.org/Team:UMaryland/project/futureapplications"><span class="yjm_has_none"><span class="yjm_title">future applications</span></span></a></span></li><li class="item301 level1 lilast"><span class="mymarg"><a class="yjanchor last" href="https://igem.org/Safety/Safety_Form?team_id=1489"><span class="yjm_has_none"><span class="yjm_title">Safety</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li><li class="haschild item234 level0"><span class="child"><a class="yjanchor " href="#"><span class="yjm_has_none"><span class="yjm_title">Notebook</span></span></a></span><ul class="subul_main level1"><li class="bl"></li><li class="tl"></li><li class="tr"></li><li class="item303 level1 first"><span class="mymarg"><a class="yjanchor first" href="https://2014.igem.org/Team:UMaryland/notebook"><span class="yjm_has_none"><span class="yjm_title">Wetlab Notebook</span></span></a></span></li><li class="item294 level1 lilast"><a class="yjanchor last" href="https://2014.igem.org/Team:UMaryland/notebook" target="_blank" ><span class="yjm_has_none"><span class="yjm_title">Protocols</span></span></a></li><li class="right"></li><li class="br"></li></ul></li><li class="haschild item123 level0"><span class="child"><a class="yjanchor " href="#"><span class="yjm_has_none"><span class="yjm_title">Human Practices</span></span></a></span><ul class="subul_main level1"><li class="bl"></li><li class="tl"></li><li class="tr"></li><li class="item106 level1 first"><span class="mymarg"><a class="yjanchor first" href="https://2014.igem.org/Team:UMaryland/humanprac/overview"><span class="yjm_has_none"><span class="yjm_title">Overview | Mission Statement</span></span></a></span></li><li class="haschild item129 level1"><span class="child subparent"><a class="yjanchor grouptitle" href="https://2014.igem.org/Team:UMaryland/humanprac/overview"><span class="yjm_has_none"><span class="yjm_title">Public School Outreach</span></span></a></span><ul class="subul_main level2"><li class="bl"></li><li class="tl"></li><li class="tr"></li><li class="item134 level2 first lilast"><span class="mymarg"><a class="yjanchor firstlast" href="https://2014.igem.org/Team:UMaryland/humanprac/publicschool"><span class="yjm_has_none"><span class="yjm_title">HighSchool</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li><li class="item131 level1"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/humanprac/community"><span class="yjm_has_none"><span class="yjm_title">Community Interaction</span></span></a></span></li><li class="item132 level1 lilast"><span class="mymarg"><a class="yjanchor last" href="https://2014.igem.org/Team:UMaryland/humanprac/publicawareness"><span class="yjm_has_none"><span class="yjm_title">Public Awareness</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li></ul></div><br />
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<h1>Meet our awesome team</h1><br />
<p> </p><br />
<p>Find our more about the three groups of contributors that made our project possible</p></div></div></div></div> </div><br />
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<br />
<br />
<br />
<div class="teaserarticles multicolumns"><br />
<div class="first float-left width33"><br />
<br />
<div class="news_item_c teamPage"><br />
<h2 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/meetourteam/tanya"><br />
Tanya Gordonov </a><br />
</h2><br />
<div class="clr"></div><br />
<div class="newsitem_text"><br />
<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/1/1d/TanyaGordonov.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong>Graduate Advisor|Bioengineering</strong></p><br />
<p> </p><br />
</div><br />
<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/tanya"><br />
<span><br />
Read more: Tanya Gordonov </span><br />
</a><br />
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<span class="article_separator">&nbsp;</span><br />
</div><br />
<br />
<br />
<div class="float-left width33"><br />
<br />
<div class="news_item_c teamPage"><br />
<h2 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/meetourteam/adip"><br />
Adip Bhargav </a><br />
</h2><br />
<div class="clr"></div><br />
<div class="newsitem_text"><br />
<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/1/13/AdipBhargav.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate|Sophomore|Neurobiology &amp; Physiology</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
<p> </p><br />
</div><br />
<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/adip"><br />
<span><br />
Read more: Adip Bhargav </span><br />
</a><br />
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<br />
<div class="news_item_c teamPage"><br />
<h2 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/meetourteam/aditya"><br />
Aditya Biswas </a><br />
</h2><br />
<div class="clr"></div><br />
<div class="newsitem_text"><br />
<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/6/6d/Adityabiswas.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate|Junior|Bioengineering</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
<p> </p><br />
</div><br />
<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/aditya"><br />
<span><br />
Read more: Aditya Biswas </span><br />
</a><br />
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<span class="article_separator">&nbsp;</span><br />
</div><br />
<br />
<span class="row-separator"></span><br />
<br />
<div class="first float-left width33"><br />
<br />
<div class="news_item_c teamPage"><br />
<h2 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/meetourteam/navadeep"><br />
Navadeep Boruah </a><br />
</h2><br />
<div class="clr"></div><br />
<div class="newsitem_text"><br />
<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/0/08/Navadeep2.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong>Graduate Advisor|Chemical and Biomolecular Engineering</strong></p><br />
<p> </p><br />
</div><br />
<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/navadeep"><br />
<span><br />
Read more: Navadeep Boruah </span><br />
</a><br />
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<span class="article_separator">&nbsp;</span><br />
</div><br />
<br />
<br />
<div class="float-left width33"><br />
<br />
<div class="news_item_c teamPage"><br />
<h2 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/meetourteam/andrew"><br />
Andrew Zhao </a><br />
</h2><br />
<div class="clr"></div><br />
<div class="newsitem_text"><br />
<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/0/06/AndrewZhao.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate|Sophomore|Bioengineering</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
<p> </p><br />
</div><br />
<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/andrew"><br />
<span><br />
Read more: Andrew Zhao </span><br />
</a><br />
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<br />
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<br />
<div class="news_item_c teamPage"><br />
<h2 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/meetourteam/kevin"><br />
Kevin Pineault </a><br />
</h2><br />
<div class="clr"></div><br />
<div class="newsitem_text"><br />
<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/4/42/KevinPineault.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong>Undergraduate|Junior|Bioengineering</strong></p><br />
<p> </p><br />
</div><br />
<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/kevin"><br />
<span><br />
Read more: Kevin Pineault </span><br />
</a><br />
</div><br />
<span class="article_separator">&nbsp;</span><br />
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<span class="row-separator"></span><br />
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<div class="first float-left width33"><br />
<br />
<div class="news_item_c teamPage"><br />
<h2 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/meetourteam/iowis"><br />
Iowis Zhu </a><br />
</h2><br />
<div class="clr"></div><br />
<div class="newsitem_text"><br />
<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/d/d1/IowisZhu.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate|Junior|BioChemistry/Cell Biology and Molecular Genetics|Statistics Minor</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
<p> </p><br />
</div><br />
<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/iowis"><br />
<span><br />
Read more: Iowis Zhu </span><br />
</a><br />
</div><br />
<span class="article_separator">&nbsp;</span><br />
</div><br />
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<br />
<div class="float-left width33"><br />
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<div class="news_item_c teamPage"><br />
<h2 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/meetourteam/chris"><br />
Chris Euerle </a><br />
</h2><br />
<div class="clr"></div><br />
<div class="newsitem_text"><br />
<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/d/d6/Chriseuerle.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate|Junior|Computer Science/ Nuerobiology &amp; Physiology</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
<p> </p><br />
</div><br />
<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/chris"><br />
<span><br />
Read more: Chris Euerle </span><br />
</a><br />
</div><br />
<span class="article_separator">&nbsp;</span><br />
</div><br />
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<div class="last float-left width33"><br />
<br />
<div class="news_item_c teamPage"><br />
<h2 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/meetourteam/boots"><br />
Dr. Boots Quimby </a><br />
</h2><br />
<div class="clr"></div><br />
<div class="newsitem_text"><br />
<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/f/f0/Bootsquimby.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong>Faculty Advisor|Integrated Life Sciences</strong></p><br />
<p> </p><br />
</div><br />
<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/boots"><br />
<span><br />
Read more: Dr. Boots Quimby </span><br />
</a><br />
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<div class="news_item_c teamPage"><br />
<h2 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/meetourteam/janine"><br />
Janine Taira </a><br />
</h2><br />
<div class="clr"></div><br />
<div class="newsitem_text"><br />
<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/4/49/JanineTaira.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate|Sophomore|Neurobiology &amp; Physiology</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
<p> </p><br />
</div><br />
<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/janine"><br />
<span><br />
Read more: Janine Taira </span><br />
</a><br />
</div><br />
<span class="article_separator">&nbsp;</span><br />
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<div class="float-left width33"><br />
<br />
<div class="news_item_c teamPage"><br />
<h2 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/meetourteam/camila"><br />
Camila Uechi </a><br />
</h2><br />
<div class="clr"></div><br />
<div class="newsitem_text"><br />
<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/6/63/Camilauechi.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate|Junior|Bioengineering</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
<p> </p><br />
</div><br />
<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/camila"><br />
<span><br />
Read more: Camila Uechi </span><br />
</a><br />
</div><br />
<span class="article_separator">&nbsp;</span><br />
</div><br />
<br />
<br />
<div class="last float-left width33"><br />
<br />
<div class="news_item_c teamPage"><br />
<h2 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/meetourteam/jason"><br />
Jason Kahn </a><br />
</h2><br />
<div class="clr"></div><br />
<div class="newsitem_text"><br />
<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/d/d1/JasonKahn.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong>Faculty Advisor|Biochemistry</strong></p><br />
<p> </p><br />
</div><br />
<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/jason"><br />
<span><br />
Read more: Jason Kahn </span><br />
</a><br />
</div><br />
<span class="article_separator">&nbsp;</span><br />
</div><br />
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<span class="row-separator"></span><br />
<br />
<div class="first float-left width33"><br />
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<div class="news_item_c teamPage"><br />
<h2 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/meetourteam/nathan"><br />
Nathan Barber </a><br />
</h2><br />
<div class="clr"></div><br />
<div class="newsitem_text"><br />
<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/8/81/NathanBarber.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate|Senior|Bioengineering|Engineering Leadership Minor</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
<p> </p><br />
</div><br />
<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/nathan"><br />
<span><br />
Read more: Nathan Barber </span><br />
</a><br />
</div><br />
<span class="article_separator">&nbsp;</span><br />
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<br />
<br />
<div class="float-left width33"><br />
<br />
<div class="news_item_c teamPage"><br />
<h2 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/meetourteam/pavel"><br />
Pavel Alexandrovich Vlasov </a><br />
</h2><br />
<div class="clr"></div><br />
<div class="newsitem_text"><br />
<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/5/55/Pavelvlasov.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate|Sophomore|Biochemistry/Cell Biology &amp; Molecular Genetics</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
<p> </p><br />
</div><br />
<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/pavel"><br />
<span><br />
Read more: Pavel Alexandrovich Vlasov </span><br />
</a><br />
</div><br />
<span class="article_separator">&nbsp;</span><br />
</div><br />
<br />
<br />
<div class="last float-left width33"><br />
<br />
<div class="news_item_c teamPage"><br />
<h2 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/meetourteam/ed"><br />
Dr. Edward Eisenstein </a><br />
</h2><br />
<div class="clr"></div><br />
<div class="newsitem_text"><br />
<p><img class="teamImage" src="https://static.igem.org/mediawiki/parts/7/77/Edeisenstein.jpg" border="0" alt="team john" width="208" height="208" style="border: 0;" /></p><br />
<p> </p><br />
<p style="text-align: center;"><strong>Faculty Advisor|Bioengineering</strong></p><br />
<p> </p><br />
</div><br />
<a class="btn btn-small btn-sm readon" href="https://2014.igem.org/Team:UMaryland/meetourteam/ed"><br />
<span><br />
Read more: Dr. Edward Eisenstein </span><br />
</a><br />
</div><br />
<span class="article_separator">&nbsp;</span><br />
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<div class="yjsg-morearticles jbsm"><br />
<h3>Subcategories</h3><br />
<br />
<ul class="categories_view"><br />
<li class="first"><br />
<span class="catitem_title"><a href="https://2014.igem.org/Team:UMaryland/meetourteam/grad"><br />
Graduate Student Advisors</a><br />
</span><br />
<br />
<br />
<br />
</li><br />
<li><br />
<span class="catitem_title"><a href="https://2014.igem.org/Team:UMaryland/meetourteam/undergrad"><br />
Undergraduate Students</a><br />
</span><br />
<br />
<br />
<br />
</li><br />
<li class="last"><br />
<span class="catitem_title"><a href="https://2014.igem.org/Team:UMaryland/meetourteam/faculty"><br />
Faculty Advisors</a><br />
</span><br />
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</li><br />
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<div class="yjsg7_before"><div id="yjsg7" class="yjsg_grid yjsgsitew"><div id="user23" class="yjsgxhtml only_mod"><div class="yjsquare modid117"><div class="h2_holder"><h2 class="module_title"><span class="title_split titlesplit0">About</span> <span class="title_split titlesplit1">Umaryland</span></h2></div><div class="yjsquare_in"><p>UMaryland2014 is University of Maryland, College Parks, inaugural iGEM team. We are a combined effort of several departments and numerous faculty mentors. Although it is only our first year, believe our hard work and dedication has paid off. We can't wait for this years competition! GO TERPS!</p></div></div></div></div></div> <div class="footer_holders footer"><!-- footer --><br />
<br />
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<script type="text/javascript"> var logo_w = '240'; var site_w = '1200'; var site_f = '13px'; var sp='https://2014.igem.org/Team:UMaryland'; var tp ='baseline'; var compileme =0; var fontc ='baseline_73331408039633'; var bootstrapv='bootstrap2'; var yver='2'; var yjsglegacy='1'; var yjsgrtl='2'; var menuanimation='fade';var menuanimationspeed=300; var YJSG_topmenu_font = '13px'; (function($){ $(window).load(function(){ $('.horiznav').SmoothDropJQ({ contpoz:0, horizLeftOffset: 20, horizRightOffset: -20, horizTopOffset: 20, verticalTopOffset:30, verticalLeftOffset: 10, maxOutside: 50 }); }) })(jQuery); </script><br />
</div> <div id="mmenu_holder"><br />
<span class="yjmm_select" id="yjmm_selectid">Meet Our Team Members</span><br />
<select id="mmenu" class="yjstyled"><br />
<option value="https://2014.igem.org/Team:UMaryland">&nbsp;Homepage</option><br />
<option value="https://2014.igem.org/Team:UMaryland/photos">&nbsp;Photos</option><br />
<option value="https://2014.igem.org/Team:UMaryland/meetourteam">&nbsp;Team</option><br />
<option value="https://2014.igem.org/Team:UMaryland/meetourteam" selected="selected">&nbsp;--Meet Our Team Members</option><br />
<option value="https://igem.org/Team.cgi?id=1489">&nbsp;--iGEM Page</option><br />
<option value="https://2014.igem.org/Team:UMaryland/team/attributions">&nbsp;--Attributions</option><br />
<option value="https://2014.igem.org/Team:UMaryland/team/sponsors">&nbsp;--Sponsors</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/overview">&nbsp;Project</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/overview">&nbsp;--Overview</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/background">&nbsp;--Background</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/biobricks">&nbsp;--parts (BioBricks)</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/results">&nbsp;--Results</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/futureapplications">&nbsp;--future applications</option><br />
<option value="https://igem.org/Safety/Safety_Form?team_id=1489">&nbsp;--Safety</option><br />
<option value="https://2014.igem.org/Team:UMaryland/notebook">&nbsp;Notebook</option><br />
<option value="https://2014.igem.org/Team:UMaryland/notebook">&nbsp;--Wetlab Notebook</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/overview">&nbsp;Human Practices</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/overview">&nbsp;--Overview | Mission Statement</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicschool">&nbsp;--Public School Outreach</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicschool">&nbsp;---HighSchool</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/community">&nbsp;--Community Interaction</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicawareness">&nbsp;--Public Awareness</option><br />
</select><br />
</div> </body><br />
</html></div>
Nbb826
http://2014.igem.org/Team:UMaryland/photos
Team:UMaryland/photos
2014-10-16T22:11:59Z
<p>Nbb826: </p>
<hr />
<div>{{UMaryland/CSS}}<br />
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<p><span style="line-height: 12.0000009536743px;">Chesapeake Bay Foundation Visit!</span></p><br />
<p><img src="https://static.igem.org/mediawiki/parts/a/a6/Bayfoundation1.jpg" border="0" width="300" height="234" style="border: 0;" /></p><br />
<p><span style="font-size: 11.0pt; line-height: 115%; font-family: 'Calibri','sans-serif'; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-hansi-theme-font: minor-latin; mso-bidi-font-family: 'Times New Roman'; mso-bidi-theme-font: minor-bidi; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;">Iowis Zhu and Sampada Koshatwar worked at the Chesapeake Bay Foundation’s Oyster Restoration/Oyster Gardening Center. </span></p><br />
<p><span style="font-family: Calibri, sans-serif; font-size: 14.4444446563721px; line-height: 16.8666667938232px;">Oyster Farm Visit</span></p><br />
<p><img src="https://static.igem.org/mediawiki/parts/8/8d/Oysterfarm.jpg" border="0" width="300" height="212" style="border: 0;" /></p><br />
<p> <span style="font-family: Calibri, sans-serif; font-size: 11pt; line-height: 115%;">From left: Iowis Zhu, Janine Taira, oyster farmer Kevin McClaren, Cami Uechi, and Adip Bhargav. The team spoke with an oyster farmer to learn more about the different perspectives surrounding the dermo infestation levels in the Chesapeake.</span></p><br />
<p> </p><br />
<p><span style="font-size: 11.0pt; line-height: 115%; font-family: 'Calibri','sans-serif'; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-hansi-theme-font: minor-latin; mso-bidi-font-family: 'Times New Roman'; mso-bidi-theme-font: minor-bidi; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;"> <img src="https://static.igem.org/mediawiki/parts/c/c8/Oysterfarm2.jpg" border="0" width="286" height="161" style="border: 0;" /></span><img src="https://static.igem.org/mediawiki/parts/5/53/Oysterfarm3.jpg" border="0" width="288" height="163" style="font-family: Calibri, sans-serif; font-size: 14.4444446563721px; line-height: 16.8666667938232px; cursor: se-resize !important; border: 0;" /></p><br />
<p><img src="https://static.igem.org/mediawiki/parts/6/65/Oysterfarm4.jpg" border="0" width="289" height="164" style="border: 0;" /></p><br />
<p>Churchill School Visit</p><br />
<p><img src="https://static.igem.org/mediawiki/parts/b/bb/Pavelschoolvisit.jpg" border="0" width="289" height="216" style="border: 0;" /></p><br />
<p><span style="font-size: 11.0pt; line-height: 115%; font-family: 'Calibri','sans-serif'; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-hansi-theme-font: minor-latin; mso-bidi-font-family: 'Times New Roman'; mso-bidi-theme-font: minor-bidi; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;">Junior Pavel Vlasov presented the project to students at Winston Churchill High School to foster interest in synthetic biology.</span></p><br />
<p><span style="font-size: 11.0pt; line-height: 115%; font-family: 'Calibri','sans-serif'; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-hansi-theme-font: minor-latin; mso-bidi-font-family: 'Times New Roman'; mso-bidi-theme-font: minor-bidi; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;"> </span></p><br />
<p>Colloboration Visits</p><br />
<p><img src="https://static.igem.org/mediawiki/parts/8/80/UmarylandVastavisit.jpg" border="0" width="291" height="163" /></p><br />
<p><span style="font-size: 11.0pt; line-height: 115%; font-family: 'Calibri','sans-serif'; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-hansi-theme-font: minor-latin; mso-bidi-font-family: 'Times New Roman'; mso-bidi-theme-font: minor-bidi; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;">From left: Dr. Gerardo Vasta, Janine Taira, Adip Bhargav, Iowis Zhu, and Andrew Zhao. The UMaryland iGem team met with Dr. Vasta, who discovered the CvGal1 receptor.</span></p><br />
<p><span style="font-size: 11.0pt; line-height: 115%; font-family: 'Calibri','sans-serif'; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-hansi-theme-font: minor-latin; mso-bidi-font-family: 'Times New Roman'; mso-bidi-theme-font: minor-bidi; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;"> </span></p><br />
<p>Visit to the DNR</p><br />
<p> <img src="https://static.igem.org/mediawiki/parts/a/a6/UMDNR1.jpg" border="0" width="226" height="169" /></p><br />
<p><img src="https://static.igem.org/mediawiki/parts/1/15/UMDNR2.jpg" border="0" width="219" height="292" /></p><br />
<p><img src="https://static.igem.org/mediawiki/parts/2/22/UMDNR3.jpg" border="0" width="219" height="164" /></p><br />
<p> </p><br />
<p><img src="https://static.igem.org/mediawiki/parts/9/99/UMDNR4.jpg" border="0" width="216" height="287" /></p><br />
<p><img src="https://static.igem.org/mediawiki/parts/c/c7/UMDNR5.jpg" border="0" width="214" height="286" /></p><br />
<p>Janine Taira, Iowis Zhu, Aditya Biswas and Andrew Zhao all visited the Maryland Department of Natural Resources to learn more about Dermo, including current laboratory detection methods.</p> </div> <br />
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<option value="https://2014.igem.org/Team:UMaryland">&nbsp;Homepage</option><br />
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<option value="https://2014.igem.org/Team:UMaryland/meetourteam">&nbsp;Team</option><br />
<option value="https://2014.igem.org/Team:UMaryland/meetourteam">&nbsp;--Meet Our Team Members</option><br />
<option value="https://igem.org/Team.cgi?id=1489">&nbsp;--iGEM Page</option><br />
<option value="https://2014.igem.org/Team:UMaryland/team/attributions">&nbsp;--Attributions</option><br />
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<option value="https://2014.igem.org/Team:UMaryland/project/biobricks">&nbsp;--parts (BioBricks)</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/results">&nbsp;--Results</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/futureapplications">&nbsp;--future applications</option><br />
<option value="https://igem.org/Safety/Safety_Form?team_id=1489">&nbsp;--Safety</option><br />
<option value="#">&nbsp;Notebook</option><br />
<option value="https://2014.igem.org/Team:UMaryland/notebook">&nbsp;--Wetlab Notebook</option><br />
<option value="#">&nbsp;Human Practices</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/overview">&nbsp;--Overview | Mission Statement</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicschool">&nbsp;--Public School Outreach</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicschool">&nbsp;---HighSchool</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/community">&nbsp;--Community Interaction</option><br />
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Nbb826
http://2014.igem.org/Team:UMaryland
Team:UMaryland
2014-10-16T22:10:18Z
<p>Nbb826: </p>
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first"><span class="mymarg"><a class="yjanchor first" href="https://2014.igem.org/Team:UMaryland/project/overview"><span class="yjm_has_none"><span class="yjm_title">Overview</span></span></a></span></li><li class="item145 level1"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/project/background"><span class="yjm_has_none"><span class="yjm_title">Background</span></span></a></span></li><li class="item144 level1"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/project/biobricks"><span class="yjm_has_none"><span class="yjm_title">parts (BioBricks)</span></span></a></span></li><li class="item102 level1"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/project/results" target="_blank" ><span class="yjm_has_none"><span class="yjm_title">Results</span></span></a></li><li class="item254 level1"><span class="mymarg"><a class="yjanchor " 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<p>Home of The University of Maryland's first iGEM team.</p><br />
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<p><a href="https://2014.igem.org/Team:UMaryland/project/overview" target="_self"><img src="https://static.igem.org/mediawiki/parts/5/55/UMIgem_logo_%281%29.png" border="0" alt="Our research heavily focuses on saving the Chesapeake Bay through protecting the oysters" width="1000" height="364" onmouseover="this.src='https://static.igem.org/mediawiki/parts/f/f2/Umarylandigeminfographic.jpg'" onmouseout="this.src= 'https://static.igem.org/mediawiki/parts/5/55/UMIgem_logo_%281%29.png'" style="border: 0;" /></a></p></div></div></div></div> </div><br />
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<p><a href="https://2014.igem.org/Team:UMaryland/meetourteam" target="_self"><img src="https://static.igem.org/mediawiki/parts/7/7a/Umarylandlabphoto.jpg" border="0" alt="Our research heavily focuses on saving the Chesapeake Bay through protecting the oysters" width="500" height="150" onmouseover="this.src='https://static.igem.org/mediawiki/parts/d/d4/TeamPhoto.JPG'" onmouseout="this.src= 'https://static.igem.org/mediawiki/parts/7/7a/Umarylandlabphoto.jpg'" style="border: 0;" /></a></p></div></div></div></div></div> <div class="yjsg7_before"><div id="yjsg7" class="yjsg_grid yjsgsitew"><div id="user23" class="yjsgxhtml only_mod"><div class="yjsquare modid117"><div class="h2_holder"><h2 class="module_title"><span class="title_split titlesplit0">About</span> <span class="title_split titlesplit1">Umaryland</span></h2></div><div class="yjsquare_in"><p>UMaryland2014 is University of Maryland, College Parks, inaugural iGEM team. We are a combined effort of several departments and numerous faculty mentors. Although it is only our first year, believe our hard work and dedication has paid off. We can't wait for this years competition! GO TERPS!</p></div></div></div></div></div> <div class="footer_holders footer"><!-- footer --><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/project/biobricks
Team:UMaryland/project/biobricks
2014-10-16T22:09:05Z
<p>Nbb826: </p>
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<h1 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/project/biobricks " class="contentpagetitle"> <br />
Biobricks <br />
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<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; text-decoration: underline;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">Part:BBa_K1489002 | </span></span><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap; text-decoration: underline;">Lpp-OmpA in new backbone</span></strong></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/parts/f/fd/PSB1C3_-_BBa_K1489002.png" border="0" width="480" height="335" style="border: 0;" /></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;">We moved an existing biobrick, BBa_K103006, from an old plasmid backbone, pSB1A2, into the current backbone, pSB1C3, via RE cloning.</span></p><br />
<p style="text-align: center;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 20px; font-weight: bold; line-height: 19.5px; text-align: start;">Lpp-OmpA-Linker (pSB1C3)</span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">Outer membrane protein A (OmpA) is a common outer membrane protein found in bacteria that serves various purposes. For our purposes, the beta-barrel transmembrane domain can be utilized to anchor otherwise soluble proteins in the outer cell membrane. Lpp serves to direct the fusion protein, while a unstructured linker (GGGSGGGS) serves to separate OmpA from its cargo in order to avoid interactions between the two proteins.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">The expressed protein is structurally identical to BBa_K103006. This version of the biobrick has been moved into the new backbone, pSB1C3, which contains the gene for chloramphenicol resistance. This resistance gene contains an internal SacI site, making the SacI site at the end of the gene almost unusable for RE cloning. BBa_K1489003 mutagenizes this SacI site into a KasI in order to avoid this issue.</p><br />
<p><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;"> </span></p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="line-height: 12.0000009536743px; color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; text-decoration: underline;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">Part:BBa_K1489000 | </span></span><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap; text-decoration: underline;">pBAD-RBS-Bovine Galectin 1-B0012 Terminator</span></strong></span></p><br />
<p><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/parts/4/42/PSB1C3_-_BBa_K1489000.png" border="0" width="515" height="335" style="border: 0px; display: block; margin-left: auto; margin-right: auto;" /></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;">A soluble his-tagged galectin from Bos taurus, expressed under Arabinose induction</span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px; text-align: center;"><span style="font-size: 20px; font-weight: bold;">pBAD-RBS-Bovine Galectin-1-B0012 Terminator</span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">Soluble beta-galactoside binding protein (galectin) originally from Bos taurus. It is believed to expressed both intracellularly and extracellularly, where they serve to binding various types of beta-galactosides and initialize signal cascades. It is believed to be a dimer in its active form, as is true with most mammalian galectins.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">This biobrick is under the pBAD promoter (BBa_K206000) with attached RBS (BBa_B0034), and utilizes an additional RNA polymerase terminator (BBa_B0012).</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">UMaryland 2014 is interested in utilizing this part to investigate whether E. coli can recognize and bind to surface carbohydrates on a marine pathogen. Other uses of this part lie in the recognition of various carbohydrate ligands and potential activation of signal transduction pathways.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;"> </p><br />
<p style="text-align: center;"> </p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="text-align: left; line-height: 17.9400005340576px; white-space: pre-wrap;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; text-decoration: underline;">Part:BBa_K1489004 | </span></span><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; line-height: 17.9400005340576px; white-space: pre-wrap; text-decoration: underline;">pBAD-RBS-Lpp-OmpA-Bovine Galectin 1-B0012 Terminator</span></strong></span></p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; line-height: 17.9400005340576px; white-space: pre-wrap; text-decoration: underline;"><img src="https://static.igem.org/mediawiki/parts/0/0f/PSB1C3_-_BBa_K1489004.png" border="0" width="515" height="335" /></span></strong></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">His-tagged bovine galectin 1 attached to the c-terminus of OmpA, expressed under arabinose induction</span></span></p><br />
<p> </p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 20px; font-weight: bold; line-height: 19.5px; text-align: start;">pBAD-RBS-OmpA-Bovine Galectin-1-B0012 Terminator</span></span></p><br />
<p style="text-align: left;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 13px; line-height: 19.5px; text-align: start;">A fusion protein of OmpA (BBa_K1489002) fused to Bovine Galectin-1 (BBa_K1489001). Used to bind sugars at the outer cell membrane. Under pBAD regulation.</span></span></p><br />
<p style="text-align: left;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"> </span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"><span style="text-decoration: underline;"><strong>Part:BBa_K1489003 | Revised Lpp-OmpA</strong></span></span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"><span style="text-decoration: underline;"><strong><img src="https://static.igem.org/mediawiki/parts/9/90/PSB1C3_-_BBa_K1489003.png" border="0" width="515" height="335" /></strong></span></span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"> Biobrick 1, with a SacI site at the C-terminus replaced by a KasI site. This improvement was made in order to re-enable RE cloning; a SacI site exists in the chloramphenicol resistance gene.</span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 20px; font-weight: bold; line-height: 19.5px; text-align: start;">Lpp-OmpA-Linker with KasI site</span></span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">Outer membrane protein A (OmpA) is a common outer membrane protein found in bacteria that serves various purposes. For our purposes, the beta-barrel transmembrane domain can be utilized to anchor otherwise soluble proteins in the outer cell membrane. Lpp serves to direct the fusion protein, while a unstructured linker (GGGSGGGS) serves to separate OmpA from its cargo in order to avoid interactions between the two proteins.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">The expressed protein is almost structurally identical to BBa_K103006, with the terminal serine on the linker replaced by an alanine. This change was made in order to convert the previous SacI site (GAGCTC), which was unusable for RE cloning due to a second SacI site being present in the plasmid backbone, into a KasI site (GGCGCC). The KasI site codes for Gly-Ala, while the SacI site codes for Gly-Ser. This amino acid substitution is unlikely to impact the function of Lpp-OmpA-Linker in a major way.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;"> </p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;"> </p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">Part:BBa_K1489001 | Bovine Galectin-1</span></span></strong></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/parts/4/4c/PSB1C3_-_BBa_K1489001.png" border="0" width="515" height="335" style="border: 0;" /></span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">This is the Bovine Galectin which should interact with beta-galactosides and intialize a signal cascase</span></span></p><br />
<p style="text-align: center;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 20px; font-weight: bold; line-height: 19.5px; text-align: start;">Bovine Galectin-1</span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">Soluble beta-galactoside binding protein (galectin) originally from Bos taurus. It is believed to expressed both intracellularly and extracellularly, where they serve to binding various types of beta-galactosides and initialize signal cascades. It is believed to be a dimer in its active form, as is true with most mammalian galectins.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">UMaryland 2014 is interested in utilizing this part to investigate whether E. coli can recognize and bind to surface carbohydrates on a marine pathogen. Other uses of this part lie in the recognition of various carbohydrate ligands and potential activation of signal transduction pathways.</p> <br />
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<option value="https://2014.igem.org/Team:UMaryland/meetourteam">&nbsp;Team</option><br />
<option value="https://2014.igem.org/Team:UMaryland/meetourteam">&nbsp;--Meet Our Team Members</option><br />
<option value="https://igem.org/Team.cgi?id=1489">&nbsp;--iGEM Page</option><br />
<option value="https://2014.igem.org/Team:UMaryland/team/attributions">&nbsp;--Attributions</option><br />
<option value="https://2014.igem.org/Team:UMaryland/team/sponsors">&nbsp;--Sponsors</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/overview">&nbsp;Project</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/overview">&nbsp;--Overview</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/background">&nbsp;--Background</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/biobricks " selected="selected">&nbsp;--parts (BioBricks)</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/results">&nbsp;--Results</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/futureapplications">&nbsp;--future applications</option><br />
<option value="https://igem.org/Safety/Safety_Form?team_id=1489">&nbsp;--Safety</option><br />
<option value="https://2014.igem.org/Team:UMaryland/notebook">&nbsp;Notebook</option><br />
<option value="https://2014.igem.org/Team:UMaryland/notebook">&nbsp;--Wetlab Notebook</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/overview">&nbsp;Human Practices</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/overview">&nbsp;--Overview | Mission Statement</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicschool">&nbsp;--Public School Outreach</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicschool">&nbsp;---HighSchool</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/community">&nbsp;--Community Interaction</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicawareness">&nbsp;--Public Awareness</option><br />
</select><br />
</div> </body><br />
</html></div>
Nbb826
http://2014.igem.org/Team:UMaryland/project/biobricks
Team:UMaryland/project/biobricks
2014-10-16T22:05:10Z
<p>Nbb826: </p>
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<h1 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/project/biobricks " class="contentpagetitle"> <br />
Biobricks <br />
</a><br />
</h1><br />
<br />
<br />
<br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; text-decoration: underline;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">Part:BBa_K1489002 | </span></span><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap; text-decoration: underline;">Lpp-OmpA in new backbone</span></strong></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/parts/f/fd/PSB1C3_-_BBa_K1489002.png" border="0" width="480" height="335" style="border: 0;" /></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;">We moved an existing biobrick, BBa_K103006, from an old plasmid backbone, pSB1A2, into the current backbone, pSB1C3, via RE cloning.</span></p><br />
<p style="text-align: center;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 20px; font-weight: bold; line-height: 19.5px; text-align: start;">Lpp-OmpA-Linker (pSB1C3)</span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">Outer membrane protein A (OmpA) is a common outer membrane protein found in bacteria that serves various purposes. For our purposes, the beta-barrel transmembrane domain can be utilized to anchor otherwise soluble proteins in the outer cell membrane. Lpp serves to direct the fusion protein, while a unstructured linker (GGGSGGGS) serves to separate OmpA from its cargo in order to avoid interactions between the two proteins.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">The expressed protein is structurally identical to BBa_K103006. This version of the biobrick has been moved into the new backbone, pSB1C3, which contains the gene for chloramphenicol resistance. This resistance gene contains an internal SacI site, making the SacI site at the end of the gene almost unusable for RE cloning. BBa_K1489003 mutagenizes this SacI site into a KasI in order to avoid this issue.</p><br />
<p><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;"> </span></p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="line-height: 12.0000009536743px; color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; text-decoration: underline;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">Part:BBa_K1489000 | </span></span><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap; text-decoration: underline;">pBAD-RBS-Bovine Galectin 1-B0012 Terminator</span></strong></span></p><br />
<p><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/parts/4/42/PSB1C3_-_BBa_K1489000.png" border="0" width="515" height="335" style="border: 0px; display: block; margin-left: auto; margin-right: auto;" /></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;">A soluble his-tagged galectin from Bos taurus, expressed under Arabinose induction</span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px; text-align: center;"><span style="font-size: 20px; font-weight: bold;">pBAD-RBS-Bovine Galectin-1-B0012 Terminator</span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">Soluble beta-galactoside binding protein (galectin) originally from Bos taurus. It is believed to expressed both intracellularly and extracellularly, where they serve to binding various types of beta-galactosides and initialize signal cascades. It is believed to be a dimer in its active form, as is true with most mammalian galectins.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">This biobrick is under the pBAD promoter (BBa_K206000) with attached RBS (BBa_B0034), and utilizes an additional RNA polymerase terminator (BBa_B0012).</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">UMaryland 2014 is interested in utilizing this part to investigate whether E. coli can recognize and bind to surface carbohydrates on a marine pathogen. Other uses of this part lie in the recognition of various carbohydrate ligands and potential activation of signal transduction pathways.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;"> </p><br />
<p style="text-align: center;"> </p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="text-align: left; line-height: 17.9400005340576px; white-space: pre-wrap;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; text-decoration: underline;">Part:BBa_K1489004 | </span></span><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; line-height: 17.9400005340576px; white-space: pre-wrap; text-decoration: underline;">pBAD-RBS-Lpp-OmpA-Bovine Galectin 1-B0012 Terminator</span></strong></span></p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; line-height: 17.9400005340576px; white-space: pre-wrap; text-decoration: underline;"><img src="https://static.igem.org/mediawiki/parts/0/0f/PSB1C3_-_BBa_K1489004.png" border="0" width="515" height="335" /></span></strong></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">His-tagged bovine galectin 1 attached to the c-terminus of OmpA, expressed under arabinose induction</span></span></p><br />
<p> </p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 20px; font-weight: bold; line-height: 19.5px; text-align: start;">pBAD-RBS-OmpA-Bovine Galectin-1-B0012 Terminator</span></span></p><br />
<p style="text-align: left;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 13px; line-height: 19.5px; text-align: start;">A fusion protein of OmpA (BBa_K1489002) fused to Bovine Galectin-1 (BBa_K1489001). Used to bind sugars at the outer cell membrane. Under pBAD regulation.</span></span></p><br />
<p style="text-align: left;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"> </span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"><span style="text-decoration: underline;"><strong>Part:BBa_K1489003 | Revised Lpp-OmpA</strong></span></span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"><span style="text-decoration: underline;"><strong><img src="https://static.igem.org/mediawiki/parts/9/90/PSB1C3_-_BBa_K1489003.png" border="0" width="515" height="335" /></strong></span></span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"> Biobrick 1, with a SacI site at the C-terminus replaced by a KasI site. This improvement was made in order to re-enable RE cloning; a SacI site exists in the chloramphenicol resistance gene.</span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 20px; font-weight: bold; line-height: 19.5px; text-align: start;">Lpp-OmpA-Linker with KasI site</span></span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">Outer membrane protein A (OmpA) is a common outer membrane protein found in bacteria that serves various purposes. For our purposes, the beta-barrel transmembrane domain can be utilized to anchor otherwise soluble proteins in the outer cell membrane. Lpp serves to direct the fusion protein, while a unstructured linker (GGGSGGGS) serves to separate OmpA from its cargo in order to avoid interactions between the two proteins.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">The expressed protein is almost structurally identical to BBa_K103006, with the terminal serine on the linker replaced by an alanine. This change was made in order to convert the previous SacI site (GAGCTC), which was unusable for RE cloning due to a second SacI site being present in the plasmid backbone, into a KasI site (GGCGCC). The KasI site codes for Gly-Ala, while the SacI site codes for Gly-Ser. This amino acid substitution is unlikely to impact the function of Lpp-OmpA-Linker in a major way.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;"> </p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;"> </p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">Part:BBa_K1489001 | Bovine Galectin-1</span></span></strong></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/parts/4/4c/PSB1C3_-_BBa_K1489001.png" border="0" width="515" height="335" style="border: 0;" /></span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">This is the Bovine Galectin which should interact with beta-galactosides and intialize a signal cascase</span></span></p><br />
<p style="text-align: center;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 20px; font-weight: bold; line-height: 19.5px; text-align: start;">Bovine Galectin-1</span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">Soluble beta-galactoside binding protein (galectin) originally from Bos taurus. It is believed to expressed both intracellularly and extracellularly, where they serve to binding various types of beta-galactosides and initialize signal cascades. It is believed to be a dimer in its active form, as is true with most mammalian galectins.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">UMaryland 2014 is interested in utilizing this part to investigate whether E. coli can recognize and bind to surface carbohydrates on a marine pathogen. Other uses of this part lie in the recognition of various carbohydrate ligands and potential activation of signal transduction pathways.</p> <br />
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Team:UMaryland/humanprac/publicschool
2014-10-16T21:50:54Z
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School Outreach <br />
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<p> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-align: center;" dir="ltr"><span style="text-decoration: underline;"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: underline; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">Churchill High School</span></strong></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="text-decoration: underline;"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: underline; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;"> </span></strong></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="text-decoration: underline;"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: underline; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;"> </span></strong></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-align: center;" dir="ltr"><span style="text-decoration: underline;"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: underline; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;"><img src="https://static.igem.org/mediawiki/parts/b/bb/Pavelschoolvisit.jpg" border="0" width="494" height="371" style="border: 0;" /><br /></span></strong></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">In June of 2014, a member of our group conducted a short presentation about genetic engineering for an AP Biology class at Winston Churchill High School in Potomac, Maryland. The presentation contained information on the process of genetic engineering in general including transcription and translation, the basics of bacterial genetics, plasmid design; and on our team’s project including the importance of oysters to the Chesapeake Bay ecosystem and economy, the mechanism by which </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> infects oysters, and the protein structure being designed for our project. Approximately half of the class period, both during the presentation and after it, was left for students to ask questions. Some of the students were clearly interested in learning more about the current and future applications of genetic engineering, such as in agriculture, and about the possible risks involved and how they can be reduced. Some of the students asking these questions later said after the presentation that they were interested in studying genetics in college. Overall, the presentation was understandable and engaging to most of the class and several students were clearly interested in learning more about IGEM and our team’s current project.</span></p> <br />
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<div class="yjsg7_before"><div id="yjsg7" class="yjsg_grid yjsgsitew"><div id="user23" class="yjsgxhtml only_mod"><div class="yjsquare modid117"><div class="h2_holder"><h2 class="module_title"><span class="title_split titlesplit0">About</span> <span class="title_split titlesplit1">Umaryland</span></h2></div><div class="yjsquare_in"><p>UMaryland2014 is University of Maryland, College Parks, inaugural iGEM team. We are a combined effort of several departments and numerous faculty mentors. Although it is only our first year, believe our hard work and dedication has paid off. We can't wait for this years competition! GO TERPS!</p></div></div></div></div></div> <div class="footer_holders footer"><!-- footer --><br />
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<option value="https://2014.igem.org/Team:UMaryland/project/futureapplications">&nbsp;--future applications</option><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/humanprac/publicawareness
Team:UMaryland/humanprac/publicawareness
2014-10-16T21:49:49Z
<p>Nbb826: </p>
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first"><span class="mymarg"><a class="yjanchor first" href="https://2014.igem.org/Team:UMaryland/project/overview"><span class="yjm_has_none"><span class="yjm_title">Overview</span></span></a></span></li><li class="item145 level1"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/project/background"><span class="yjm_has_none"><span class="yjm_title">Background</span></span></a></span></li><li class="item144 level1"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/project/biobricks"><span class="yjm_has_none"><span class="yjm_title">parts (BioBricks)</span></span></a></span></li><li class="item102 level1"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/project/results" target="_blank" ><span class="yjm_has_none"><span class="yjm_title">Results</span></span></a></li><li class="item254 level1"><span class="mymarg"><a class="yjanchor " 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class="right"></li><li class="br"></li></ul></li><li class=" active haschild item123 level0"><span class="child"><a class="yjanchor activepath " href="https://2014.igem.org/Team:UMaryland/humanprac/overview"><span class="yjm_has_none"><span class="yjm_title">Human Practices</span></span></a></span><ul class="subul_main level1"><li class="bl"></li><li class="tl"></li><li class="tr"></li><li class="item106 level1 first"><span class="mymarg"><a class="yjanchor first" href="https://2014.igem.org/Team:UMaryland/humanprac/overview"><span class="yjm_has_none"><span class="yjm_title">Overview | Mission Statement</span></span></a></span></li><li class="haschild item129 level1"><span class="child subparent"><a class="yjanchor grouptitle" href="https://2014.igem.org/Team:UMaryland/humanprac/publicschool"><span class="yjm_has_none"><span class="yjm_title">Public School Outreach</span></span></a></span><ul class="subul_main level2"><li class="bl"></li><li class="tl"></li><li class="tr"></li><li class="item134 level2 first lilast"><span class="mymarg"><a class="yjanchor firstlast" href="https://2014.igem.org/Team:UMaryland/humanprac/publicschool"><span class="yjm_has_none"><span class="yjm_title">HighSchool</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li><li class="item131 level1"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/humanprac/community"><span class="yjm_has_none"><span class="yjm_title">Community Interaction</span></span></a></span></li><li id="current" class=" active item132 level1 lilast"><span class="mymarg"><a class="yjanchor last activepath " href="https://2014.igem.org/Team:UMaryland/humanprac/publicawareness"><span class="yjm_has_none"><span class="yjm_title">Public Awareness</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li></ul></div><br />
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<h1 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/humanprac/publicawareness" class="contentpagetitle"> <br />
Public Awareness <br />
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<p class="MsoNormal" style="text-align: center;"><strong><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"><span style="text-decoration: underline;">Public Awareness</span></span></strong></p><br />
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<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">Realizing the importance of education and understanding, we strived to improve general understanding of <em>P. marinus</em> and the Dermo infection. We began at the University of Maryland with a survey to get students from all backgrounds to start thinking about the problems we were addressing. Next, we publicized our project and goals by connecting with deans of various colleges to spark interest in these fields. As awareness increased on campus, we turned our attention to the surrounding community. Networking with schools around the area, programs were established where iGEM members would go to the school and give a presentation or seminar tailored to the interests of the audience. For example, we gave a seminar on biotechnology and synthetic biology at a local high school in order to start education early with students who will be at the forefront of this type of research in the near future. The next step will be to get in touch with state organizations such as the Chesapeake Bay foundation in order to devise outreach initiatives and progressively raise awareness at higher levels of the community.</span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">Additionally we conducted a survey of over 250 students in undergraduate microbiology classes at the University of Maryland to further understand the public perception of this technology through the eyes of our next generation.The survey and responses are found below.</span></p><br />
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<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"> </span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">All responses to the survey will be kept anonymous, and no personal information about the participants will be released. The results of the survey may be published.</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;"> </span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">I have read and consent to the terms of confidentiality listed above ___________</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">1) What is a biosensor?</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">a. an organ that measures biological phenomenon</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">b. an organism that has the ability to sense other organisms</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">c. a device that measures biological parameters such as temperature or pressure</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><strong><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">d. an instrument that has a biological component and measures physical or chemical parameters</span></strong></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><strong><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;"> </span></strong></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">2) How do you feel about genetic engineering?</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">a. Strongly Support</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">b. Support</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">c. Neutral</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">d. Oppose</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">e. Strongly Oppose</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;"> </span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">3) How do you feel about the introduction of genetically modified </span><em style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">E. Coli</em><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;"> into the Chesapeake Bay?</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">a. Strongly Support</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">b. Support</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">c. Neutral</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">d. Oppose</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">e. Strongly Oppose</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;"> </span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">4) If it improved Chesapeake bay health and aquaculture?</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">a. Strongly Support</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">b. Support</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">c. Neutral</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">d. Oppose</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">e. Strongly Oppose</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;"> </span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">5) What danger do you think lies in using biotechnology and bioengineering to address environmental issues?</span></p><br />
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<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">(open ended)</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; text-align: center; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="text-decoration: underline;"><strong><span style="font-size: 10pt; font-family: Arial, sans-serif; color: #222222; text-decoration: underline;">ANSWER</span></strong></span><strong><span style="font-size: 10pt; font-family: Arial, sans-serif; color: #222222; text-decoration: underline;">S</span></strong></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;"><img src="https://static.igem.org/mediawiki/2014/1/15/UMSurveyAnswer1.jpg" border="0" width="365" height="268" style="float: left;" /></span><strong style="text-align: center;"><span style="font-size: 10pt; font-family: Arial, sans-serif; color: #222222; text-decoration: underline;"><img src="https://static.igem.org/mediawiki/2014/e/e9/UMSurveyAnswer2.jpg" border="0" width="410" height="268" style="cursor: se-resize !important; border: 0;" /></span></strong></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"> </p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><img src="https://static.igem.org/mediawiki/2014/0/07/UMSurveyAnswer3.jpg" border="0" width="365" height="268" style="border: 0;" /><img src="https://static.igem.org/mediawiki/2014/d/d2/UMSurveyAnswer4.jpg" border="0" width="365" height="268" /></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">By administering this survey to an undergraduate general microbiology class at the University of Maryland, we hoped to gain insight about where students stood on genetic engineering and its applications. We chose this class because it is a general education class at the University of Maryland which attracts students of various backgrounds and majors, not just science majors.</span></p><br />
<p class="MsoNormal"><span style="font-family: 'Times New Roman', serif; font-size: 12pt; line-height: 115%;"> With the first question, we tried to figure out just what proportion of the students were science majors or non-science majors, so that we could better interpret the responses to the later questions. We chose to ask what a biosensor was to determine if the student had significant background in biology or perhaps exposure to biotechnology. To our surprise, most students answered this question correctly which suggests that many students probably had some experience with or have heard about aspects of bioengineering. This conclusion reveals the extent of Science, Technology, Engineering, and Mathematics (STEM) education of University of Maryland students and the fact that most are fairly well informed about this type of technology.</span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">The next three questions corroborate the results from first question as shown by a relatively high number of students who are open to the idea of genetic engineering of <em>E. coli</em>. Even after being asked bluntly about the introduction of genetically-altered <em>E. coli</em> into the bay without any other information, more students, surprisingly, remained neutral rather than opposed. Finally, as expected, after qualifying the previous question by providing the motivation for the use of <em>E. coli</em>, a large shift was observed towards being supportive of the proposition. </span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"> Lastly, the free response questions (responses not shown) provided a deeper insight into the understanding of the population being surveyed. Most of the students developed valid points regarding possible areas of concern with this type of project. These ranged from ethical concerns to long-term, unexpected problems and the idea that native species may be adversely affected by our biosensor. It seemed to us that most of the concerns stemmed from a lack of understanding of the technology being used. Education and outreach efforts would likely eliminate some of these misunderstandings; however, some of the concerns raised are valid and are points we considered ourselves.</span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">The next step in this project would be to broaden the population being surveyed both in terms of size and diversity. In this way, we can determine if the results we obtained here is actually representative of a university setting. In terms of the participants being surveyed, we believe that young students are the most important because this is the generation that will be deciding on novel technologies that address pressing social and environmental concerns such as the health of the Chesapeake Bay. As a result, it is important to ensure that this demographic, especially, has access to correct information so that informed decisions can be made. However, we acknowledge that it is also important to gauge the views of current policy makers and people outside the university as well, so future surveys may be targeted at older demographics and different regions.</span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">With regards to the effect on public outreach efforts, it is clear that although most of the responses appeared to be informed there is room for improvement. Accordingly, we plan to continue our on campus and community efforts to advertise the issue and our research in order to start a thoughtful dialogue between those affected by concerns regarding the Chesapeake Bay and those who are trying to address these concerns.</span></p><br />
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<option value="https://2014.igem.org/Team:UMaryland/meetourteam">&nbsp;--Meet Our Team Members</option><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/notebook
Team:UMaryland/notebook
2014-10-16T21:36:04Z
<p>Nbb826: </p>
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<h1 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/notebook" class="contentpagetitle"> <br />
Notebook <br />
</a><br />
</h1><br />
<br />
<br />
<p class="MsoNormal"><strong>February to May </strong></p><br />
<p class="MsoNormal">Faculty and students interested in participating in UMaryland iGEM began meeting in February. Smaller interest meetings had occurred before to spread the word, as this is our first year! In these meetings, ideas were thrown around and brainstormed, and the logistics of participating in the iGEM competition were discussed. Every week for the first few months, a different student would give a presentation on a project and wiki from a previous year of iGEM in order to give all members a feel for what sort of projects have been done and what to expect throughout the process. In the later months, students presented then their own ideas for what UMaryland iGEM should pursue in 2014. Near the end of the semester, the group finally decided which of the proposed projects we would take – the development of a biosensor which would detect for the eastern oyster pathogen, <em>P. marinus</em>, in real-time using CvGal1, a galactoside-binding protein (galectin) found in the hemocytes of oysters, which has a strong affinity for binding to <em>P. marinus</em>.</p><br />
<p class="MsoNormal"> </p><br />
<p class="MsoNormal"><strong>May</strong></p><br />
<p class="MsoNormal">UMaryland iGEM began working in the lab at the end of May. Having chosen the outer membrane protein A (OmpA) to express the galectin on the surface of bacteria, the team ordered the part (BBa_K103006) and transformed it into DHα <em>E. coli</em> to create a stock of this plasmid. Team members also learned and refreshed on standard lab practices and protocols which would carry throughout most of the project: chemical transformation and bacterial culture, aseptic technique, minipreparation of a plasmid, and gel electrophoresis. From gel electrophoresis of the miniprep samples, cut at the unique BioBrick prefix and suffix with restriction enzymes EcoRI and PstI, as well as sequencing done by GENEWIZ, Inc., we could confirm that the OmpA plasmid was successfully cloned. This procedure would become our standard for cloning and confirming the identity of all subsequent plasmids.</p><br />
<p class="MsoNormal"> </p><br />
<p class="MsoNormal"><strong>June</strong></p><br />
<p class="MsoNormal">The team also began mapping out the BioBrick constructs intended for this year.</p><br />
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<p class="MsoNormal">The two constructs presented above differ only by the promoter sequence: the pBAD promoter is activated by arabinose, while the pLAC promoter is activated by IPTG. For this reason, the team also prepared a stock of pSB1C3-pBAD-RBS-GFP (BBa_K750000) and pSB1C3-pLAC-RBS-RFP (BBa_J04450) plasmids. By removing the fluorescent protein genes and linearizing the DNA, we would use these vectors as the basis for all the clones we intended to express, as the promoter, ribosomal binding site, and terminator are already part of the BioBrick pSB1C3 backbone.</p><br />
<p class="MsoNormal">Appropriately-designed primers were ordered from Integrated DNA Technologies (IDT) for preparation of Gibson assembly. With the primers, we PCR amplified the OmpA gene from its plasmid and the two vectors mentioned previously. Gels confirmed the unique sizes of the PCR products, and a NanoDrop spectrophotometer analyzed the DNA concentrations. The use of a NanoDrop machine would also become a standard in our DNA analysis, particularly for Gibson assembly, where concentrations are vital.</p><br />
<p class="MsoNormal">Due to issues with acquiring the CvGal1 gene, the team began a side project in the meantime. β-galactosidase, readily available as a BioBrick, was considered in place of CvGal1. Instead of binding a galactose sugar, as the galectin would, β-galactosidase would instead cleave it into monosaccharides. The LacZα (BBa_K329005) and full-length LacZ (BBa_K1155007) genes were prepared similar to OmpA, with the end result being PCR products of the isolated, desired LacZ gene. Since we were working with mainly DH5α <em>E. coli</em>, which have the LacZΩ fragment already present, the LacZα fragment expressed in DH5α is expected to combine and express the whole β-galactosidase. This mechanism was the preferred method, as the LacZα fragment is only around 240 bp in size. The full-length LacZ gene (approx. 3 kbp) was also used, however, in the event that the two-fragment method did not work.</p><br />
<p class="MsoNormal">The team also moved the OmpA gene from its original backone, the old pSB1A2 (ampicillin resistance), to the current BioBrick standard, pSB1C3 (chloramphenicol resistance). Using standard restriction enzyme cloning techniques and the pSB1C3 linearized backbone supplied by iGEM, the team was able to “cut-and-paste” the OmpA gene into its new vector. The process was fairly simple, due to the different antibiotic resistances acting as a general screen between the old and new plasmids. This was our first BioBrick made (BBa_K1489002), classifying as an improvement to an existing part.</p><br />
<p class="MsoNormal"> </p><br />
<p class="MsoNormal"><strong>July</strong></p><br />
<p class="MsoNormal">Going into July, the team continued this LacZ side project. We encountered many roadblocks when attempting to clone the LacZ gene into the pSB1C3-pBAD-RBS vector (pBAD was eventually chosen as the promoter of choice for the rest of our project) via Gibson reactions. The only viable plasmid produced, screened through blue colonies on X-gal-coated plate, was an attempt at cloning OmpA-LacZα. However, both the gel and sequencing results revealed a serious error within the OmpA gene.</p><br />
<p class="MsoNormal">Around the same time, at the end of July, we had received both CvGal1 and bovine galectin-1 gene fragments, ordered from IDT’s gBlock system. Both gBlocks were based on the wildtype genes, but modified to ensure a proper codon reading frame and to remove potential problems with the expressed protein, such as disulfide bonds forming due to the oxidative nature of the extracellular matrix. Immediately, the team PCR amplified all the fragments for Gibson assembly, where we faced an unresponsive CvGal1 gene. Every other fragment looked correct on the gel, however, so the team proceeded with focusing solely on bovine galectin-1 while troubleshooting CvGal1.</p><br />
<p class="MsoNormal"> </p><br />
<p class="MsoNormal"><strong>August</strong></p><br />
<p class="MsoNormal">August was filled with many trials, but two main goals: 1) Gibson cloning of the bovine galectin gene into the pSB1C3-pBAD-RBS backbone, one with fusion to the OmpA gene and one without, and 2) PCR amplification of the CvGal1 gBlock.</p><br />
<p class="MsoNormal">For the first goal, the team faced many issues with residual plasmid transforming, making it unclear whether the Gibson reaction was assembling the gene fragments properly. Finally, on August 15, it seemed we had at last cloned the bovine galectin into the vector (without OmpA). (Though done in parallel, the OmpA-Bovine galectin construct did not clone properly.) The gel revealed the plasmid’s identity clearly, but for the wrong reason – the bovine galectin gene had an EcoRI site in the middle of its sequence which had gone unnoticed, until this product was cloned and analyzed.</p><br />
<p class="MsoNormal">As for the CvGal1 gBlock, after various PCR attempts in which the team tested different Mg<sup>2+</sup> concentrations, DMSO concentrations, cycle time and temperature conditions, and DNA polymerases, the team contacted IDT about the inability to amplify the gene fragment. To the time of this writing, they are still in the process of remaking the CvGal1 gBlock, as the persistent issue appeared to be on their end.</p><br />
<p class="MsoNormal"> </p><br />
<p class="MsoNormal"><strong>September</strong></p><br />
<p class="MsoNormal">With the school semester starting again, our team members had much less time to spend in the lab. However, we continued to make progress, and toward the end of September, we had successfully cloned the OmpA-Bovine galectin construct as well. Persistence, fine-tuned procedures, and a bit of luck paid dividends as the sequencing results came back as a clear match.</p><br />
<p class="MsoNormal">The case of the unwanted EcoRI site remained within the bovine galectin genes, but the team was ready with a site-directed mutagenesis kit and primers. Mutagenesis on the EcoRI site to create a silent mutation was successful on its first try, much to our relief, and a second round of sequencing revealed that the EcoRI site in the bovine galectin gene in both constructs had been removed (but the BioBrick prefix’s EcoRI site remained intact).</p><br />
<p> </p><br />
<p class="MsoNormal">These are our two main project BioBricks: the Bovine galectin alone in pSB1C3-pBAD-RBS (BBa_K1489000) and the Bovine galectin gene fused to the end of the OmpA gene in the same vector (BBa_K1489004). Additionally, since the pSB1C3 backbone has a SacI site (which pSB1A2 did not) and OmpA has a SacI site (as designed by the creators of that BioBrick), the pSB1C3-OmpA which we created back in June had two unusable SacI sites. In order to maintain the integrity of that BioBrick with its unique restriction sites (along with the standard EcoRI and PstI sites), we also performed SDM on the pSB1C3-OmpA (BBa_K1489002) to replace the now-useless SacI site at the end of the OmpA gene with a KasI site, creating our fourth BioBrick (BBa_K1489003).</p><br />
<p class="MsoNormal"><strong>October</strong></p><br />
<p class="MsoNormal">With the BioBricks made, the team began preliminary protein expression tests. Multiple cultures of DH5α E. coli, transformed with either the bovine galectin or OmpA-BvGal1 BioBricks, were grown up to a noticeable OD. They were then induced with arabinose at increasing concentrations, from no induction to 0.2% m/v. After further incubation of the bacteria to allow for protein production, a 1 mL sample of each culture was pelleted. The cells were then lysed by resuspension in 20 μL SDS and boiling the solution, yielding a soluble protein fraction in the supernatant after centrifugation. The remaining pellet was boiled in 100 μL SDS with harsher conditions in order to extract as much insoluble protein as possible. These protein samples (soluble and insoluble fractions) were run through SDS-PAGE, followed by western blotting (taking advantage of the His6 tag placed in the synthesized bovine galectin-1). Anti-mouse was used for the secondary antibody. Expression of the bovine galectin BioBrick (BBa_K1489000) appeared strong and at the correct size, but expression of OmpA-BvGal1 (BBa_K1489004) featured weak bands in both the induced and uninduced samples, though of correct size.</p> <br />
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http://2014.igem.org/Team:UMaryland/meetourteam/janine
Team:UMaryland/meetourteam/janine
2014-10-16T21:10:30Z
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Janine Taira <br />
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<p style="text-align: center;"><strong style="color: #646464; font-family: Arial, Helvetica, sans-serif; font-size: 13px; line-height: 21.0340023040772px;">Undergraduate|Sophomore|Neurobiology &amp; Physiology</strong><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px; text-align: left;"><br /></span></p><br />
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<h4 class="yjtboxtitle">Janine Taira:</h4><br />
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<p><span style="color: #000000; font-family: sans-serif; font-size: 13px; line-height: 19.0499992370605px;">I'm planning on going to dental school after my undergraduate degree. I'm participating iGem because I want to have an hands-on experience in the lab, and modifying bacteria for competition seemed like an interesting and unique way to get that experience. It's super cool that in less than a year, we'll have some kind of a product to show for all of our work. iGem is my first research experience and has been an invaluable part of my education.</span></p><br />
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http://2014.igem.org/Team:UMaryland/humanprac/publicawareness
Team:UMaryland/humanprac/publicawareness
2014-10-16T21:08:54Z
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Publicawareness <br />
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<p class="MsoNormal" style="text-align: center;"><strong><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"><span style="text-decoration: underline;">Public Awareness</span></span></strong></p><br />
<p> </p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">Realizing the importance of education and understanding, we strived to improve general understanding of <em>P. marinus</em> and the Dermo infection. We began at the University of Maryland with a survey to get students from all backgrounds to start thinking about the problems we were addressing. Next, we publicized our project and goals by connecting with deans of various colleges to spark interest in these fields. As awareness increased on campus, we turned our attention to the surrounding community. Networking with schools around the area, programs were established where iGEM members would go to the school and give a presentation or seminar tailored to the interests of the audience. For example, we gave a seminar on biotechnology and synthetic biology at a local high school in order to start education early with students who will be at the forefront of this type of research in the near future. The next step will be to get in touch with state organizations such as the Chesapeake Bay foundation in order to devise outreach initiatives and progressively raise awareness at higher levels of the community.</span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">Additionally we conducted a survey of over 250 students in undergraduate microbiology classes at the University of Maryland to further understand the public perception of this technology through the eyes of our next generation.The survey and responses are found below.</span></p><br />
<p class="MsoNormal"> </p><br />
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<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"> </span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">All responses to the survey will be kept anonymous, and no personal information about the participants will be released. The results of the survey may be published.</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;"> </span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">I have read and consent to the terms of confidentiality listed above ___________</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">1) What is a biosensor?</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">a. an organ that measures biological phenomenon</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">b. an organism that has the ability to sense other organisms</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">c. a device that measures biological parameters such as temperature or pressure</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><strong><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">d. an instrument that has a biological component and measures physical or chemical parameters</span></strong></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><strong><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;"> </span></strong></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">2) How do you feel about genetic engineering?</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">a. Strongly Support</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">b. Support</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">c. Neutral</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">d. Oppose</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">e. Strongly Oppose</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;"> </span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">3) How do you feel about the introduction of genetically modified </span><em style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">E. Coli</em><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;"> into the Chesapeake Bay?</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">a. Strongly Support</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">b. Support</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">c. Neutral</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">d. Oppose</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">e. Strongly Oppose</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;"> </span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">4) If it improved Chesapeake bay health and aquaculture?</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">a. Strongly Support</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="color: #222222; font-family: Arial, sans-serif; font-size: 10pt;">b. Support</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">c. Neutral</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">d. Oppose</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">e. Strongly Oppose</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;"> </span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">5) What danger do you think lies in using biotechnology and bioengineering to address environmental issues?</span></p><br />
<p class="MsoNormal"> </p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;">(open ended)</span></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; text-align: center; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="text-decoration: underline;"><strong><span style="font-size: 10pt; font-family: Arial, sans-serif; color: #222222; text-decoration: underline;">ANSWER</span></strong></span><strong><span style="font-size: 10pt; font-family: Arial, sans-serif; color: #222222; text-decoration: underline;">S</span></strong></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><span style="font-size: 10.0pt; font-family: 'Arial','sans-serif'; mso-fareast-font-family: 'Times New Roman'; color: #222222;"><img src="https://static.igem.org/mediawiki/2014/1/15/UMSurveyAnswer1.jpg" border="0" width="365" height="268" style="float: left;" /></span><strong style="text-align: center;"><span style="font-size: 10pt; font-family: Arial, sans-serif; color: #222222; text-decoration: underline;"><img src="https://static.igem.org/mediawiki/2014/e/e9/UMSurveyAnswer2.jpg" border="0" width="410" height="268" style="cursor: se-resize !important; border: 0;" /></span></strong></p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"> </p><br />
<p class="MsoNormal" style="margin-bottom: 0.0001pt; line-height: normal; background-image: initial; background-attachment: initial; background-size: initial; background-origin: initial; background-clip: initial; background-position: initial; background-repeat: initial;"><img src="https://static.igem.org/mediawiki/2014/0/07/UMSurveyAnswer3.jpg" border="0" width="365" height="268" style="border: 0;" /><img src="https://static.igem.org/mediawiki/2014/d/d2/UMSurveyAnswer4.jpg" border="0" width="365" height="268" /></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">By administering this survey to an undergraduate general microbiology class at the University of Maryland, we hoped to gain insight about where students stood on genetic engineering and its applications. We chose this class because it is a general education class at the University of Maryland which attracts students of various backgrounds and majors, not just science majors.</span></p><br />
<p class="MsoNormal"><span style="font-family: 'Times New Roman', serif; font-size: 12pt; line-height: 115%;"> With the first question, we tried to figure out just what proportion of the students were science majors or non-science majors, so that we could better interpret the responses to the later questions. We chose to ask what a biosensor was to determine if the student had significant background in biology or perhaps exposure to biotechnology. To our surprise, most students answered this question correctly which suggests that many students probably had some experience with or have heard about aspects of bioengineering. This conclusion reveals the extent of Science, Technology, Engineering, and Mathematics (STEM) education of University of Maryland students and the fact that most are fairly well informed about this type of technology.</span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">The next three questions corroborate the results from first question as shown by a relatively high number of students who are open to the idea of genetic engineering of <em>E. coli</em>. Even after being asked bluntly about the introduction of genetically-altered <em>E. coli</em> into the bay without any other information, more students, surprisingly, remained neutral rather than opposed. Finally, as expected, after qualifying the previous question by providing the motivation for the use of <em>E. coli</em>, a large shift was observed towards being supportive of the proposition. </span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"> Lastly, the free response questions (responses not shown) provided a deeper insight into the understanding of the population being surveyed. Most of the students developed valid points regarding possible areas of concern with this type of project. These ranged from ethical concerns to long-term, unexpected problems and the idea that native species may be adversely affected by our biosensor. It seemed to us that most of the concerns stemmed from a lack of understanding of the technology being used. Education and outreach efforts would likely eliminate some of these misunderstandings; however, some of the concerns raised are valid and are points we considered ourselves.</span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">The next step in this project would be to broaden the population being surveyed both in terms of size and diversity. In this way, we can determine if the results we obtained here is actually representative of a university setting. In terms of the participants being surveyed, we believe that young students are the most important because this is the generation that will be deciding on novel technologies that address pressing social and environmental concerns such as the health of the Chesapeake Bay. As a result, it is important to ensure that this demographic, especially, has access to correct information so that informed decisions can be made. However, we acknowledge that it is also important to gauge the views of current policy makers and people outside the university as well, so future surveys may be targeted at older demographics and different regions.</span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">With regards to the effect on public outreach efforts, it is clear that although most of the responses appeared to be informed there is room for improvement. Accordingly, we plan to continue our on campus and community efforts to advertise the issue and our research in order to start a thoughtful dialogue between those affected by concerns regarding the Chesapeake Bay and those who are trying to address these concerns.</span></p><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/humanprac/community
Team:UMaryland/humanprac/community
2014-10-16T21:07:34Z
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first"><span class="mymarg"><a class="yjanchor first" href="https://2014.igem.org/Team:UMaryland/project/overview"><span class="yjm_has_none"><span class="yjm_title">Overview</span></span></a></span></li><li class="item145 level1"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/project/background"><span class="yjm_has_none"><span class="yjm_title">Background</span></span></a></span></li><li class="item144 level1"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/project/biobricks"><span class="yjm_has_none"><span class="yjm_title">parts (BioBricks)</span></span></a></span></li><li class="item102 level1"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/project/results" target="_blank" ><span class="yjm_has_none"><span class="yjm_title">Results</span></span></a></li><li class="item254 level1"><span class="mymarg"><a class="yjanchor " 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Community Interaction <br />
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<p class="MsoNormal" style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">Interaction with Community</span></strong></span></p><br />
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<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">We primarily focused our efforts on three organizations in the Chesapeake Bay community: Marinetics Choptank Oyster Farm, Department of Natural Resources, and Chesapeake Bay Foundation. Each provided insight into a different dimension of the Dermo infection problem. At Marinetics, we received an economy-centric view, where we learned a great deal about how <em>P. marinus</em> actually affects commercial oyster growth. We were surprised to find that, at least in that region of the bay, the owner of the farm experienced little to no effect from the pathogen. However, he did concede that detection systems for Dermo are not adequate and expressed that a real-time biosensor would be quite useful.</span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"> </span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"><img src="https://static.igem.org/mediawiki/parts/c/c8/Oysterfarm2.jpg" border="0" width="319" height="180" style="display: block; margin-left: auto; margin-right: auto;" /></span></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">Next, we explored <em>P. marinus</em> through an environmental perspective as we spoke to members of the Chesapeake Bay Foundation. Here, we heard what was more expected of the problem including its adverse effects on oyster populations in the Chesapeake Bay. This interaction revealed the larger impacts our research would have on bay health as well as informed us about current approaches to remedy the problem. Still, we were unsure what to make of the two seemingly opposite views presented by the oyster farm and the Chesapeake Bay Foundation.</span></p><br />
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<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">Lastly, to clarify techniques and detection methods, we arranged a meeting with staff and lab members from the Department of Natural Resources. Mr. Chris Dungan, who is in charge of <em>P. marinus</em> investigation among other pathogens, gave us a comprehensive overview of how Dermo infections are detected and handled. We also discussed different aspects of policy-making and the obstacles to introducing new technology to combat these problems. For example, Mr. Dungan reconciled the apparently opposing accounts we received from the Chesapeake Bay Foundation and the oyster farm by telling us that Dermo infection is more prevalent in certain areas of the bay and will adversely affect populations in the bay but maybe not in aquaculture because oysters would be harvested and cooked before they exhibited the pathophysiology of the disease. Moreover, he acknowledged that oyster farmers tend to pay less attention to <em>P. marinus</em> in general because it is not a human pathogen. </span></p><br />
<p> </p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">Ultimately, the fact that people tend to be apprehensive about new technologies and research that could be used to solve environmental problems motivated us to help address this issue. Since we understood just how powerful public opinion can be in promoting or labeling potential solutions prematurely as dangerous, we developed two main initiatives to promote education in these technologies and to gauge the public opinion quantitatively in our local community to understand why there is such reluctance towards new technology and the reasons behind this.</span></p> <br />
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Nbb826
http://2014.igem.org/Team:UMaryland/humanprac/publicschool
Team:UMaryland/humanprac/publicschool
2014-10-16T21:07:11Z
<p>Nbb826: </p>
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<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-align: center;" dir="ltr"><span style="text-decoration: underline;"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: underline; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">Churchill HighSchool</span></strong></span></p><br />
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<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="text-decoration: underline;"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: underline; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;"> </span></strong></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-align: center;" dir="ltr"><span style="text-decoration: underline;"><strong><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: normal; font-variant: normal; text-decoration: underline; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;"><img src="https://static.igem.org/mediawiki/parts/b/bb/Pavelschoolvisit.jpg" border="0" width="494" height="371" style="border: 0;" /><br /></span></strong></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">In June of 2014, a member of our group conducted a short presentation about genetic engineering for an AP Biology class at Winston Churchill High School in Potomac, Maryland. The presentation contained information on the process of genetic engineering in general including transcription and translation, the basics of bacterial genetics, plasmid design; and on our team’s project including the importance of oysters to the Chesapeake Bay ecosystem and economy, the mechanism by which </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> infects oysters, and the protein structure being designed for our project. Approximately half of the class period, both during the presentation and after it, was left for students to ask questions. Some of the students were clearly interested in learning more about the current and future applications of genetic engineering, such as in agriculture, and about the possible risks involved and how they can be reduced. Some of the students asking these questions later said after the presentation that they were interested in studying genetics in college. Overall, the presentation was understandable and engaging to most of the class and several students were clearly interested in learning more about IGEM and our team’s current project.</span></p> <br />
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<div class="yjsg7_before"><div id="yjsg7" class="yjsg_grid yjsgsitew"><div id="user23" class="yjsgxhtml only_mod"><div class="yjsquare modid117"><div class="h2_holder"><h2 class="module_title"><span class="title_split titlesplit0">About</span> <span class="title_split titlesplit1">Umaryland</span></h2></div><div class="yjsquare_in"><p>UMaryland2014 is University of Maryland, College Parks, inaugural iGEM team. We are a combined effort of several departments and numerous faculty mentors. Although it is only our first year, believe our hard work and dedication has paid off. We can't wait for this years competition! GO TERPS!</p></div></div></div></div></div> <div class="footer_holders footer"><!-- footer --><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/humanprac/overview
Team:UMaryland/humanprac/overview
2014-10-16T21:06:53Z
<p>Nbb826: </p>
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Overviewmissionstatement <br />
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<p class="MsoNormal" style="text-align: center;"><strong><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"><span style="text-decoration: underline;">Overview Mission Statement</span></span></strong></p><br />
<p class="MsoNormal"><strong><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';"> <img src="https://static.igem.org/mediawiki/parts/6/6c/Humanpracmain.png" border="0" width="593" height="553" style="display: block; margin-left: auto; margin-right: auto;" /></span></strong></p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">From the beginning, our team wanted to pursue a project that would represent the spirit of Maryland. What better than to turn to the Chesapeake Bay, a natural resource that is priceless to Marylandians. The Chesapeake Bay, in addition to being a beautiful attraction, supports aquaculture and ecosystems that are essential to the economy and the environment. So, after choosing to address an oyster pathogen, one of many issues that threaten Chesapeake Bay health, we sought to understand the implications of our project and how it could be applied.</span></p><br />
<p> </p><br />
<p class="MsoNormal"><span style="font-size: 12.0pt; mso-bidi-font-size: 11.0pt; line-height: 115%; font-family: 'Times New Roman','serif';">We organized various meetings with different Chesapeake Bay organizations to gauge the usefulness of our research and to further optimize our design. In doing so, we were made aware of the fact that public opinion played a large part in governing what research would be admissible and what the final product of the research could look like. Then, we took it to the next step to start our own initiatives to help educate and spread awareness about synthetic biology, and iGEM specifically in the context of the Chesapeake Bay and the environment. This outreach was accomplished through surveying a population of students and giving a class for middle school students.</span></p> <br />
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<div class="yjsg7_before"><div id="yjsg7" class="yjsg_grid yjsgsitew"><div id="user23" class="yjsgxhtml only_mod"><div class="yjsquare modid117"><div class="h2_holder"><h2 class="module_title"><span class="title_split titlesplit0">About</span> <span class="title_split titlesplit1">Umaryland</span></h2></div><div class="yjsquare_in"><p>UMaryland2014 is University of Maryland, College Parks, inaugural iGEM team. We are a combined effort of several departments and numerous faculty mentors. Although it is only our first year, believe our hard work and dedication has paid off. We can't wait for this years competition! GO TERPS!</p></div></div></div></div></div> <div class="footer_holders footer"><!-- footer --><br />
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<span class="yjmm_select" id="yjmm_selectid">Overview | Mission Statement</span><br />
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<option value="/iGemWiki/">&nbsp;Homepage</option><br />
<option value="https://2014.igem.org/Team:UMaryland/photos">&nbsp;Photos</option><br />
<option value="https://2014.igem.org/Team:UMaryland/meetourteam">&nbsp;Team</option><br />
<option value="https://2014.igem.org/Team:UMaryland/meetourteam">&nbsp;--Meet Our Team Members</option><br />
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<option value="https://2014.igem.org/Team:UMaryland/project/overview">&nbsp;--Overview</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/background">&nbsp;--Background</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/biobricks">&nbsp;--parts (BioBricks)</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/results">&nbsp;--Results</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/futureapplications">&nbsp;--future applications</option><br />
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<option value="https://2014.igem.org/Team:UMaryland/notebook">&nbsp;Notebook</option><br />
<option value="https://2014.igem.org/Team:UMaryland/notebook">&nbsp;--Wetlab Notebook</option><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/notebook
Team:UMaryland/notebook
2014-10-16T21:06:13Z
<p>Nbb826: </p>
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<h1 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/notebook" class="contentpagetitle"> <br />
Notebook <br />
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</h1><br />
<br />
<br />
<p class="MsoNormal"><strong>February to May </strong></p><br />
<p class="MsoNormal">Faculty and students interested in participating in UMaryland iGEM began meeting in February. Smaller interest meetings had occurred before to spread the word, as this is our first year! In these meetings, ideas were thrown around and brainstormed, and the logistics of participating in the iGEM competition were discussed. Every week for the first few months, a different student would give a presentation on a project and wiki from a previous year of iGEM in order to give all members a feel for what sort of projects have been done and what to expect throughout the process. In the later months, students presented then their own ideas for what UMaryland iGEM should pursue in 2014. Near the end of the semester, the group finally decided which of the proposed projects we would take – the development of a biosensor which would detect for the eastern oyster pathogen, <em>P. marinus</em>, in real-time using CvGal1, a galactoside-binding protein (galectin) found in the hemocytes of oysters, which has a strong affinity for binding to <em>P. marinus</em>.</p><br />
<p class="MsoNormal"> </p><br />
<p class="MsoNormal"><strong>May</strong></p><br />
<p class="MsoNormal">UMaryland iGEM began working in the lab at the end of May. Having chosen the outer membrane protein A (OmpA) to express the galectin on the surface of bacteria, the team ordered the part (BBa_K103006) and transformed it into DHα <em>E. coli</em> to create a stock of this plasmid. Team members also learned and refreshed on standard lab practices and protocols which would carry throughout most of the project: chemical transformation and bacterial culture, aseptic technique, minipreparation of a plasmid, and gel electrophoresis. From gel electrophoresis of the miniprep samples, cut at the unique BioBrick prefix and suffix with restriction enzymes EcoRI and PstI, as well as sequencing done by GENEWIZ, Inc., we could confirm that the OmpA plasmid was successfully cloned. This procedure would become our standard for cloning and confirming the identity of all subsequent plasmids.</p><br />
<p class="MsoNormal"> </p><br />
<p class="MsoNormal"><strong>June</strong></p><br />
<p class="MsoNormal">The team also began mapping out the BioBrick constructs intended for this year.</p><br />
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<p class="MsoNormal">The two constructs presented above differ only by the promoter sequence: the pBAD promoter is activated by arabinose, while the pLAC promoter is activated by IPTG. For this reason, the team also prepared a stock of pSB1C3-pBAD-RBS-GFP (BBa_K750000) and pSB1C3-pLAC-RBS-RFP (BBa_J04450) plasmids. By removing the fluorescent protein genes and linearizing the DNA, we would use these vectors as the basis for all the clones we intended to express, as the promoter, ribosomal binding site, and terminator are already part of the BioBrick pSB1C3 backbone.</p><br />
<p class="MsoNormal">Appropriately-designed primers were ordered from Integrated DNA Technologies (IDT) for preparation of Gibson assembly. With the primers, we PCR amplified the OmpA gene from its plasmid and the two vectors mentioned previously. Gels confirmed the unique sizes of the PCR products, and a NanoDrop spectrophotometer analyzed the DNA concentrations. The use of a NanoDrop machine would also become a standard in our DNA analysis, particularly for Gibson assembly, where concentrations are vital.</p><br />
<p class="MsoNormal">Due to issues with acquiring the CvGal1 gene, the team began a side project in the meantime. β-galactosidase, readily available as a BioBrick, was considered in place of CvGal1. Instead of binding a galactose sugar, as the galectin would, β-galactosidase would instead cleave it into monosaccharides. The LacZα (BBa_K329005) and full-length LacZ (BBa_K1155007) genes were prepared similar to OmpA, with the end result being PCR products of the isolated, desired LacZ gene. Since we were working with mainly DH5α <em>E. coli</em>, which have the LacZΩ fragment already present, the LacZα fragment expressed in DH5α is expected to combine and express the whole β-galactosidase. This mechanism was the preferred method, as the LacZα fragment is only around 240 bp in size. The full-length LacZ gene (approx. 3 kbp) was also used, however, in the event that the two-fragment method did not work.</p><br />
<p class="MsoNormal">The team also moved the OmpA gene from its original backone, the old pSB1A2 (ampicillin resistance), to the current BioBrick standard, pSB1C3 (chloramphenicol resistance). Using standard restriction enzyme cloning techniques and the pSB1C3 linearized backbone supplied by iGEM, the team was able to “cut-and-paste” the OmpA gene into its new vector. The process was fairly simple, due to the different antibiotic resistances acting as a general screen between the old and new plasmids. This was our first BioBrick made (BBa_K1489002), classifying as an improvement to an existing part.</p><br />
<p class="MsoNormal"> </p><br />
<p class="MsoNormal"><strong>July</strong></p><br />
<p class="MsoNormal">Going into July, the team continued this LacZ side project. We encountered many roadblocks when attempting to clone the LacZ gene into the pSB1C3-pBAD-RBS vector (pBAD was eventually chosen as the promoter of choice for the rest of our project) via Gibson reactions. The only viable plasmid produced, screened through blue colonies on X-gal-coated plate, was an attempt at cloning OmpA-LacZα. However, both the gel and sequencing results revealed a serious error within the OmpA gene.</p><br />
<p class="MsoNormal">Around the same time, at the end of July, we had received both CvGal1 and bovine galectin-1 gene fragments, ordered from IDT’s gBlock system. Both gBlocks were based on the wildtype genes, but modified to ensure a proper codon reading frame and to remove potential problems with the expressed protein, such as disulfide bonds forming due to the oxidative nature of the extracellular matrix. Immediately, the team PCR amplified all the fragments for Gibson assembly, where we faced an unresponsive CvGal1 gene. Every other fragment looked correct on the gel, however, so the team proceeded with focusing solely on bovine galectin-1 while troubleshooting CvGal1.</p><br />
<p class="MsoNormal"> </p><br />
<p class="MsoNormal"><strong>August</strong></p><br />
<p class="MsoNormal">August was filled with many trials, but two main goals: 1) Gibson cloning of the bovine galectin gene into the pSB1C3-pBAD-RBS backbone, one with fusion to the OmpA gene and one without, and 2) PCR amplification of the CvGal1 gBlock.</p><br />
<p class="MsoNormal">For the first goal, the team faced many issues with residual plasmid transforming, making it unclear whether the Gibson reaction was assembling the gene fragments properly. Finally, on August 15, it seemed we had at last cloned the bovine galectin into the vector (without OmpA). (Though done in parallel, the OmpA-Bovine galectin construct did not clone properly.) The gel revealed the plasmid’s identity clearly, but for the wrong reason – the bovine galectin gene had an EcoRI site in the middle of its sequence which had gone unnoticed, until this product was cloned and analyzed.</p><br />
<p class="MsoNormal">As for the CvGal1 gBlock, after various PCR attempts in which the team tested different Mg<sup>2+</sup> concentrations, DMSO concentrations, cycle time and temperature conditions, and DNA polymerases, the team contacted IDT about the inability to amplify the gene fragment. To the time of this writing, they are still in the process of remaking the CvGal1 gBlock, as the persistent issue appeared to be on their end.</p><br />
<p class="MsoNormal"> </p><br />
<p class="MsoNormal"><strong>September</strong></p><br />
<p class="MsoNormal">With the school semester starting again, our team members had much less time to spend in the lab. However, we continued to make progress, and toward the end of September, we had successfully cloned the OmpA-Bovine galectin construct as well. Persistence, fine-tuned procedures, and a bit of luck paid dividends as the sequencing results came back as a clear match.</p><br />
<p class="MsoNormal">The case of the unwanted EcoRI site remained within the bovine galectin genes, but the team was ready with a site-directed mutagenesis kit and primers. Mutagenesis on the EcoRI site to create a silent mutation was successful on its first try, much to our relief, and a second round of sequencing revealed that the EcoRI site in the bovine galectin gene in both constructs had been removed (but the BioBrick prefix’s EcoRI site remained intact).</p><br />
<p> </p><br />
<p class="MsoNormal">These are our two main project BioBricks: the Bovine galectin alone in pSB1C3-pBAD-RBS (BBa_K1489000) and the Bovine galectin gene fused to the end of the OmpA gene in the same vector (BBa_K1489004). Additionally, since the pSB1C3 backbone has a SacI site (which pSB1A2 did not) and OmpA has a SacI site (as designed by the creators of that BioBrick), the pSB1C3-OmpA which we created back in June had two unusable SacI sites. In order to maintain the integrity of that BioBrick with its unique restriction sites (along with the standard EcoRI and PstI sites), we also performed SDM on the pSB1C3-OmpA (BBa_K1489002) to replace the now-useless SacI site at the end of the OmpA gene with a KasI site, creating our fourth BioBrick (BBa_K1489003).</p><br />
<p class="MsoNormal"><strong>October</strong></p><br />
<p class="MsoNormal">fillertext</p> <br />
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Future Applications <br />
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<p>Future Applications of Research</p><br />
<p>We believe that the initial stages of the Biosensor have been complete and will lead to many new and exciting developments in the future. The general idea is that once the surface protein has been expressed, it will be able to interact with Dermo and produce a signal.</p><br />
<p>As for this year we our efforts working with a Bovine-Galectin have come the furthest and are ready to move into these next steps. Additionally in future years we hope to incorporate CvGal1 into the constructs, an effort we were not able to accomplish due to lack of time and troubleshooting. </p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong>Signaling Approaches:</strong></span></p><br />
<p> </p><br />
<p>1). Genetic engineering approach -Downstream Signaling Cascade</p><br />
<p><img src="https://static.igem.org/mediawiki/parts/9/99/UMSIgnal_Cascade.jpg" border="0" width="671" height="503" style="line-height: 12.0000009536743px; display: block; margin-left: auto; margin-right: auto; cursor: se-resize !important;" /></p><br />
<p>One of the strategies we hope to pursue is developing or incorporating a downstream signaling cascade into the cell strain. In this effort, once the E. coli came in contact with the Dermo it would then set off a transcription factor, ultimately producing a signal. Multiple approaches have been researched, including a Cpx membrane stress cascade, a split GFP cascade, and others.</p><br />
<p>The Cpx two component system reacts naturally to membrane stress. When the much larger Dermo is bound to the surface receptor expressed on E. coli, the bacterial cell membrane would become deformed. In turn, this would transmit a signal to the nucleus via the Cpx system that could be coupled to GFP production. This assay would require florescence microscopy to detect Dermo in solution.</p><br />
<p>Another option is for a split GFP construct. The idea is two different constructs containing the ligand are present in the cell, each with a different fraction of the GFP gene. When Dermo comes in close contact with the cell, multiple ligands on the cell would bind to the receptors on Dermo, causing the membrane to pinch or come together. This would trigger a spontaneous chance that two complementary pieces of GFP in close contact would bind, producing a fluorescent signal. This would also need to be observed using microscopy.</p><br />
<p>One of the ultimate goals from using the downstream signaling approach is to develop a “Kamikaze Bacteria”. By this we hope that the bacteria would respond to the presence of Dermo by eliminating it, possibly through killing itself. Similar work has previously been done with bees and colony collapse.</p><br />
<p>2). Microfluidics Approach- Size Based Cell sorter</p><br />
<p style="text-align: center;"><img src="https://static.igem.org/mediawiki/parts/0/02/UMMicrofluidics.png" border="0" width="572" height="416" /></p><br />
<p>Another avenue we plan to pursue is a microfluidics based detection approach. The overall concept would rely on the fact that E.coli cells are much smaller than Dermo cells. By creating a microfluidic device that sorts based on size, E.coli would travel into one output, whereas the Dermo and Dermo+E.coli complexes would travel into another output. Furthermore if the E.coli cells are fluorescently stained, analysis of the larger size fraction would indicate whether any E.coli cells are bound, confirming the presence of Dermo. While further differentiations based on size between bound and unbound Dermo may not be possible due to growth differences, this method will still suffice. </p><br />
<p>A microfluidic device would also contain the E.coli preventing them from entering the surrounding environment. Containment would be an obvious concern if a device like this were to ever hit the market. In this solution theoretically one could have pre-labeled E.coli in the machine stored and ready. The end user would simply intake a water sample right from the source, allow it to incubate, and process (sort) it. Collecting the larger fraction and viewing under a microscope would give a quick and easy real time method for detecting Dermo.</p><br />
<p>We believe that with the groundwork we have laid this year these future endeavors will be made possible as UMaryland continues to grow and gain expertise.</p><br />
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Team:UMaryland/project/futureapplications
2014-10-16T21:02:19Z
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Future Applications <br />
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<p>Future Applications of Research</p><br />
<p>We believe that the initial stages of the Biosensor have been complete and will lead to many new and exciting developments in the future. The general idea is that once the surface protein has been expressed, it will be able to interact with Dermo and produce a signal.</p><br />
<p>As for this year we our efforts working with a Bovine-Galectin have come the furthest and are ready to move into these next steps. Additionally in future years we hope to incorporate CvGal1 into the constructs, an effort we were not able to accomplish due to lack of time and troubleshooting. </p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong>Signaling Approaches:</strong></span></p><br />
<p> </p><br />
<p>1). Genetic engineering approach -Downstream Signaling Cascade</p><br />
<p><img src="https://static.igem.org/mediawiki/parts/9/99/UMSIgnal_Cascade.jpg" border="0" width="671" height="503" style="line-height: 12.0000009536743px; display: block; margin-left: auto; margin-right: auto; cursor: se-resize !important;" /></p><br />
<p>One of the strategies we hope to pursue is developing or incorporating a downstream signaling cascade into the cell strain. In this effort, once the E. coli came in contact with the Dermo it would then set off a transcription factor, ultimately producing a signal. Multiple approaches have been researched, including a Cpx membrane stress cascade, a split GFP cascade, and others.</p><br />
<p>The Cpx two component system reacts naturally to membrane stress. When the much larger Dermo is bound to the surface receptor expressed on E. coli, the bacterial cell membrane would become deformed. In turn, this would transmit a signal to the nucleus via the Cpx system that could be coupled to GFP production. This assay would require florescence microscopy to detect Dermo in solution.</p><br />
<p>Another option is for a split GFP construct. The idea is two different constructs containing the ligand are present in the cell, each with a different fraction of the GFP gene. When Dermo comes in close contact with the cell, multiple ligands on the cell would bind to the receptors on Dermo, causing the membrane to pinch or come together. This would trigger a spontaneous chance that two complementary pieces of GFP in close contact would bind, producing a fluorescent signal. This would also need to be observed using microscopy.</p><br />
<p>One of the ultimate goals from using the downstream signaling approach is to develop a “Kamikaze Bacteria”. By this we hope that the bacteria would respond to the presence of Dermo by eliminating it, possibly through killing itself. Similar work has previously been done with bees and colony collapse.</p><br />
<p>2). Microfluidics Approach- Size Based Cell sorter</p><br />
<p style="text-align: center;"><img src="https://static.igem.org/mediawiki/parts/0/02/UMMicrofluidics.png" border="0" width="572" height="416" /></p><br />
<p>Another avenue we plan to pursue is a microfluidics based detection approach. The overall concept would rely on the fact that E.coli cells are much smaller than Dermo cells. By creating a microfluidic device that sorts based on size, E.coli would travel into one output, whereas the Dermo and Dermo+E.coli complexes would travel into another output. Furthermore if the E.coli cells are fluorescently stained, analysis of the larger size fraction would indicate whether any E.coli cells are bound, confirming the presence of Dermo. While further differentiations based on size between bound and unbound Dermo may not be possible due to growth differences, this method will still suffice. </p><br />
<p>A microfluidic device would also contain the E.coli preventing them from entering the surrounding environment. Containment would be an obvious concern if a device like this were to ever hit the market. In this solution theoretically one could have pre-labeled E.coli in the machine stored and ready. The end user would simply intake a water sample right from the source, allow it to incubate, and process (sort) it. Collecting the larger fraction and viewing under a microscope would give a quick and easy real time method for detecting Dermo.</p><br />
<p>We believe that with the groundwork we have laid this year these future endeavors will be made possible as UMaryland continues to grow and gain expertise.</p><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/project/results
Team:UMaryland/project/results
2014-10-16T21:01:50Z
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<h1 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/project/results" class="contentpagetitle"> <br />
Results <br />
</a><br />
</h1><br />
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<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; text-indent: 36pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/2014/e/e0/UMFig1-1.jpg" border="0" width="213px;" height="248px;" style="border: NaNpx solid black; transform: rotate(0.00rad); -webkit-transform: rotate(0.00rad);" /></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/2014/2/28/UMFig1-2.png" border="0" width="354px;" height="148px;" style="border: NaNpx solid black; transform: rotate(0.00rad); -webkit-transform: rotate(0.00rad);" /></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> A B</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig. 1 - Bovine Galectin-1 gene cloned into the pSB1C3 vector with pBAD promoter and a RBS</span></p><br />
<p><strong id="docs-internal-guid-809a97bc-f600-9f66-fada-73accace1d98" style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Galectin-1 from </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Bos taurus</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> has been cloned into pSB1C3 vector. </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Bos taurus </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">or bovine galectin enabled us to generate a platform that enables future work on ligand binding. While CvGals binding characteristics are still being researched the crystal structure and nature of bovine galectin has been well studied. The subsequent BioBricks regarding this gene were all constructed via Gibson assembly and cloning. The plasmids from which the OmpA insert and the vector were amplified via extension PCR are respectively: OmpA with linker (BBa_K103006) and pBAD-RBS on the pSB1C3 backbone (BBa_K750000). For the latter, the GFP in the BioBrick was not included for Gibson assembly. The bovine galectin-1 was ordered as a synthetically synthesized sequence from IDT as a linear gene and amplified directly with appropriate primers for Gibson assembly. </span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Figure </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">1A</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> shows the Bovine galectin-1 assembled into pSB1C3-pBAD-RBS alone (no OmpA), which when expressed in </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">E. coli</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> should produce a soluble form of the protein. After successful assembly and cloned into DH5α </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">E. coli</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">. The plasmid was cut with restriction enzymes EcoRI and PstI, standard sites in the pSB1C3 vector’s BioBrick prefix and suffix, respectively. The result is expected to yield the insert, pBAD-RBS-Bovine galectin-1, and the pSB1C3 vector. However, an unwanted EcoRI site was found in the galectin-1 gene afterwards, yielding the pattern seen in the gel. Figure </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">1B</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> demonstrates the expected pattern, with band sizes of 2033 bp, 496 bp, and 169 bp top to bottom, via the software “A plasmid Editor” (ApE, courtesy of M. Wayne Davis, University of Utah).</span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/2014/3/3f/UMFig2-1.jpg" border="0" width="233" height="331" style="border: 0;" />A<img src="https://static.igem.org/mediawiki/2014/e/ef/UMFig2-2.png" border="0" width="354px;" height="151px;" style="border: 0; transform: rotate(0.00rad); -webkit-transform: rotate(0.00rad);" /></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">B</span></p><br />
<p><strong style="font-weight: normal;"> (3rd Lane contains OmpA-Bovine product)</strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig. 2 - </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">OmpA-Bovine (cut with BamHI and HindIII, before SDM)</span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Figure </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">2A</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> shows the Bovine galectin-1 assembled into pSB1C3-pBAD-RBS as a fusion protein with OmpA. When expressed in </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">E. coli</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">, the galectin should continue off the C-terminus of the OmpA-linker and thus be transported to the extracellular matrix since it is attached to the OmpA transport protein. After plasmid assembly transfer of plasmid into a DH5α </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">E. coli</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> cells strain, the plasmid was extracted and digested with EcoRI and PstI. The pattern of three bands appears again due to an additional EcoRI restriction site. </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 12.0000009536743px;"><span><img src="https://static.igem.org/mediawiki/2014/4/41/UMFig4-1.jpg" border="0" width="735" height="272" /></span></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 12.0000009536743px;">Sequencing Results:</p><br />
<p style="line-height: 12.0000009536743px;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$0:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$0:0">OmpA-Bovine SDM Chromatogram:</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$1:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$2:0">Sequence file:</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$3:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$4:0">NNNNNNNNNNNTNNNNANNNNNAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGGATG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$5:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$6:0">ATTTCTGGAATTCGCGGCCGCTTCTAGAGACATTGATTATTTGCACGGCGTCACACTTTGCTATGCCATAGCAAGATAGT</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$7:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$8:0">CCATAAGATTAGCGGATCCTACCTGACGCTTTTTATCGCAACTCTCTACTGTTTCTCCATACCGTTTTTTTGGGCTAGCT</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$9:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$10:0">ACTAGAGAAAGAGGAGAAATACTAGCATATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTCTACTCTGC</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$11:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$12:0">TGGCAGGTTGCTCCAGCAACGCTAAAATCGATCAGGGAATTAACCCGTATGTTGGCTTTGAAATGGGTTACGACTGGTTA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$13:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$14:0">GGTCGTATGCCGTACAAAGGCAGCGTTGAAAACGGTGCATACAAAGCTCAGGGCGTTCAACTGACCGCTAAACTGGGTTA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$15:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$16:0">CCCAATCACTGACGACCTGGACATCTACACTCGTCTGGGTGGCATGGTATGGCGTGCAGACACTAAATCCAACGTTTATG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$17:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$18:0">GTAAAAACCACGACACCGGCGTTTCTCCGGTCTTCGCTGGCGGTGTTGAGTACGCGATCACTCCTGAAATCGCTACCCGT</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$19:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$20:0">CTGGAATACCAGTGGACCAACAACATCGGTGACGCACACACCATCGGCACTCGTCCGGACAACGGCGGANGTTCTGGAGG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$21:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$22:0">ANGGAGCATGGCGTCGGGGCTTGTTGCGTCCAATCTGAATCTGAAACCAGGTGAAAGCTTACGCGTCCGCGGCGAAGTAG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$23:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$24:0">CCGCAGATGCGAAATCTTTTCTCTTGAACCTGGGCAAGGACGANAATAATCTGGCTTTGCATTTCAACCCGCGTTTCNAC</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$25:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$26:0">GCTCATGGNGACNTGAACACAATCNTGTGTAANTCCAAGGNTGCTGGTGCTTGGGGTGCANAANNANCGNGNNNTCCGCG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$27:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$28:0">TTCCCGTTTCANNNNNGCTCANNNNGGNAGTCACCNTNCNGCNTTCANCCANNNCGANNTNCNNNNTTAAANNNGNNNNN</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$29:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$30:0">NGGNNNCNAAGTTNNANTTNNCNAANCNNNTNNAACNNNNNGNNNNNNNNNNNNNNNNNGNNNN</span></span></p><br />
<p style="line-height: 12.0000009536743px;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$0:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$30:0">A</span></span></p><br />
<p style="line-height: 12.0000009536743px;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$0:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$30:0"><img src="https://static.igem.org/mediawiki/2014/b/bc/UMFig5-1.jpg" border="0" width="733" height="272" /></span></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"> </p><br />
<p style="line-height: 12.0000009536743px;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$0:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$30:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$0:0">pBAD-Bovine SDM chromatogram:</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$1:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$2:0">NNNNNNNNNNNNTNNNCTATNNNNTAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$3:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$4:0">ATGATTTCTGGAATTCGCGGCCGCTTCTAGAGACATTGATTATTTGCACGGCGTCACACTTTGCTATGCCATAGCAAGAT</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$5:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$6:0">AGTCCATAAGATTAGCGGATCCTACCTGACGCTTTTTATCGCAACTCTCTACTGTTTCTCCATACCGTTTTTTTGGGCTA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$7:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$8:0">GCTACTAGAGAAAGAGGAGAAATACTAGATGGCGTCGGGGCTTGTTGCGTCCAATCTGAATCTGAAACCAGGTGAAAGCT</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$9:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$10:0">TACGCGTCCGCGGCGAAGTAGCCGCAGATGCGAAATCTTTTCTCTTGAACCTGGGCAAGGACGATAATAATCTGGCTTTG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$11:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$12:0">CATTTCAACCCGCGTTTCAACGCTCATGGCGACGTGAACACAATCGTGTGTAATTCCAAGGATGCTGGTGCTTGGGGTGC</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$13:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$14:0">AGAACAGCGTGAGTCCGCGTTCCCGTTTCAACCGGGCTCAGTTGTGGAAGTCACCATCAGCTTCAACCAGACCGATCTCA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$15:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$16:0">CCATTAAACTGCCGGATGGCTACGAGTTTAAGTTTCCAAACCGTTTAAACCTCGAGGCCATCAACTATCTGTCCGCGGGG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$17:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$18:0">GGCGACTTCAAGATTAAAAGCGTGCACCATCATCACCACCATTGATCACACTGGCTCACCTTCGGGTGGGCCTTTCTGCG</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$19:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$20:0">TTTATATACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTTCTTTAAAACCGA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$21:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$22:0">AAAGATTACTTCGCGTTATGCNGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$23:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$24:0">GCTCACTCAAAGGCGGTAATACGGTTATCNNCAGAATCNGGGGATNACGCAGGAAANAACATGTGAGCAAANNNCAGCAA</span><br style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$25:0" /><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$26:0">AANGCAGGAANCGTAAAANNNN</span></span></span></p><br />
<p style="line-height: 12.0000009536743px;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$0:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413325852852=213d6532e7dde259850.2:0.0.0.0.0.0.$end:0:$30:0"><span style="font-size: 12px; line-height: 15.3599996566772px;" data-reactid=".129.$mid=11413326239352=25f60c68bc4103c7554.2:0.0.0.0.0.0.$end:0:$26:0">B</span></span></span></p><br />
<p> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig. 3 - <strong>S</strong></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">equencing results and Chromatogram of SDM’d plasmids (</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">BBa_K1489000 and BBa_K1489004)</span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">To address this issue, site-directed mutagenesis was performed on these plasmids to change the EcoRI sequence within the bovine galectin-1 gene (5’-GAATTC-3’) to 5’-GAGTTT-3’, which will code for the same Glu-Phe sequence. Figure </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">3A</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> and </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">3B</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> demonstrate the two successfully mutagenized base pairs in the bovine galectin-1 gene, while preserving the rest of the plasmid.</span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p> <img src="https://static.igem.org/mediawiki/parts/a/a5/UMFinalfigureigem.jpg" border="0" width="687" height="269" style="line-height: 12.0000009536743px; cursor: se-resize !important;" /></p><br />
<p> Sequencing results:<br />
NNNNNNNNNNNNNNNNTATANAAATAGGCGTATCNCGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGG<br />
ATGATTTCTGGAATTCGCGGCCGCTTCTAGAGGGCATATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTT<br />
CTACTCTGCTGGCAGGTTGCTCCAGCAACGCTAAAATCGATCAGGGAATTAACCCGTATGTTGGCTTTGAAATGGGTTAC<br />
GACTGGTTAGGTCGTATGCCGTACAAAGGCAGCGTTGAAAACGGTGCATACAAAGCTCAGGGCGTTCAACTGACCGCTAA<br />
ACTGGGTTACCCAATCACTGACGACCTGGACATCTACACTCGTCTGGGTGGCATGGTATGGCGTGCAGACACTAAATCCA<br />
ACGTTTATGGTAAAAACCACGACACCGGCGTTTCTCCGGTCTTCGCTGGCGGTGTTGAGTACGCGATCACTCCTGAAATC<br />
GCTACCCGTCTGGAATACCAGTGGACCAACAACATCGGTGACGCACACACCATCGGCACTCGTCCGGACAACGGCGGAGG<br />
TTCTGGAGGAGGGAGCTCTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTT<br />
CTTTAAAACCGAAAAGATTACTTCGCGTTATGCAGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGG<br />
CGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCANGNNNAACATGTGAGCA<br />
AAAGGCCAGCNNAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCACAGGCTCCGCCCCCCTGACGAGCAT<br />
CACAAAAATCGACGCTCAAGTCAGANGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGNAGCTC<br />
CCTCGTGCGCTCTCCTGTTCCNACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTT<br />
CTCNTAGCTCACGCTGTAGGTATCNNCANNN</p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig 4. - <strong>S</strong></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">equencing Results and Chromatogram of OmpA moved into pSB1C3</span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Since UMaryland worked extensively with the OmpA-linker (BBa_K103006), we have also looked into improvements on that BioBrick.</span></p><br />
<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; background-color: #f7f7f7;">OmpA-KasI chromatogram, shrunk down</span></span></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 12.0000009536743px;"><img src="https://static.igem.org/mediawiki/2014/0/0b/UMFig3-1.jpg" border="0" width="687" height="269" /></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 12.0000009536743px;">Sequencing Results:</p><br />
<p style="line-height: 12.0000009536743px;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$4:0">Sequence:</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$5:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$6:0">NNNNNNNNNNNNNNNTATAAAAATAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGGA</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$7:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$8:0">TGATTTCTGGAATTCGCGGCCGCTTCTAGAGGGCATATGAAAGCTACTAAACTGGTACTGGGCGCGGTAATCCTGGGTTC</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$9:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$10:0">TACTCTGCTGGCAGGTTGCTCCAGCAACGCTAAAATCGATCAGGGAATTAACCCGTATGTTGGCTTTGAAATGGGTTACG</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$11:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$12:0">ACTGGTTAGGTCGTATGCCGTACAAAGGCAGCGTTGAAAACGGTGCATACAAAGCTCAGGGCGTTCAACTGACCGCTAAA</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$13:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$14:0">CTGGGTTACCCAATCACTGACGACCTGGACATCTACACTCGTCTGGGTGGCATGGTATGGCGTGCAGACACTAAATCCAA</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$15:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$16:0">CGTTTATGGTAAAAACCACGACACCGGCGTTTCTCCGGTCTTCGCTGGCGGTGTTGAGTACGCGATCACTCCTGAAATCG</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$17:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$18:0">CTACCCGTCTGGAATACCAGTGGACCAACAACATCGGTGACGCACACACCATCGGCACTCGTCCGGACAACGGCGGAGGT</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$19:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$20:0">TCTGGAGGAGGGGCGCCTACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCACCACCCTGCCCTTTTTC</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$21:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$22:0">TTTAAAACCGAAAAGATTACTTCGCGTTATGCAGGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGC</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$23:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$24:0">GAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCA</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$25:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$26:0">AAAGGNCAGCAAAAGGNCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCACAGGCTCCGCCCCCCTGACGAGCA</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$27:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$28:0">TCACAAAAATCGACGCTCAAGTCANNNNGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTC</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$29:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$30:0">CCTCGTGCGCTCTNCTGTTCCNACCCTGCCGCTTACCNGATACCTGTCNCNTTCTNCCTTCGGGANCGNGNGCTTTCTCA</span><br style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$31:0" /><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; white-space: pre-wrap; background-color: #f7f7f7;" data-reactid=".129.$mid=11413325660655=2d3fa5ac5f22ec0a422.2:0.0.0.0.0.0.$end:0:$32:0">TAGCTCACGCTNTAGGTANNNCANTNCGGNNNTNGNCGTTNNNCNCNANNCTNGNNTN</span></p><br />
<p> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig. 5 - <strong>S</strong></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">equencing Results and Chromatogram of SDM’d OmpA in pSB1C3</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><br /></span><span style="line-height: 12.777777671814px;">The previous version of Lpp-OmpA-Linker (BBa_K103006), designed for the pSB1A2 vector backbone, contained the non-traditional restriction sites NdeI (CATATG) and SacI (GAGCTC) in order to attach proteins at the N-terminus and C-terminus of OmpA, respectively. While the NdeI site is still useful for RE cloning at the N-terminus of OmpA, the implementation of the new plasmid backbone pSB1C3 has made the SacI site difficult to use. This is due to the presence of a second SacI site in the chloramphenicol acetyltransferase gene in pSB1C3. We have thus decided to mutagenize the previous SacI site on Lpp-OmpA-Linker into a KasI site in order to remove this issue. The KasI site (GGCGCC) encodes the amino acids glycine and alanine, converting the sequence of the linker from Gly-Gly-Gly-Ser-Gly-Gly-Gly-Ser to Gly-Gly-Gly-Ser-Gly-Gly-Gly-Ala. We believe this to be a sufficiently minor alteration to the unfolded linker sequence as to not affect protein function.</span></p> <br />
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<p><strong style="font-weight: normal;"> </strong></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><span style="color: #3e454c; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 12px; line-height: 15.3599996566772px; background-color: #f7f7f7;"></span></span></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 12.0000009536743px;"><img src="https://static.igem.org/mediawiki/parts/0/0c/UMBovinegalectinexpression.jpg" border="0" width="687" height="269" /></p><br />
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<p> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Fig. 6 - <strong>S</strong></span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Western Blot of expression of SDM’d OmpA-BovineGalectin in pSB1C3</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: transparent; font-weight: bold; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"><br /></span><span style="line-height: 12.777777671814px;">"Both the bovine galectin and the OmpA-Bovine Galectin were tested for expression using a Western blot. Each sample was also tested with and without induction. Additionally for all trials both the soluable and insoluable fractions were examined. The results showed expression of both the Bovine galectin and the OmpA-Bovine galectin. The conclusion points to sucessful expression of our biobrick "</span></p><br />
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Nbb826
http://2014.igem.org/Team:UMaryland/project/biobricks
Team:UMaryland/project/biobricks
2014-10-16T21:01:30Z
<p>Nbb826: </p>
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<div>{{UMaryland/CSS}}<br />
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<title>Parts (BioBricks)</title><br />
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<h1 class="article_title"><br />
<a href="https://2014.igem.org/Team:UMaryland/project/biobricks " class="contentpagetitle"> <br />
Biobricks <br />
</a><br />
</h1><br />
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<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; text-decoration: underline;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">Part:BBa_K1489002 | </span></span><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap; text-decoration: underline;">Lpp-OmpA in new backbone</span></strong></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/parts/f/fd/PSB1C3_-_BBa_K1489002.png" border="0" width="480" height="335" style="border: 0;" /></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;">We moved an existing biobrick, BBa_K103006, from an old plasmid backbone, pSB1A2, into the current backbone, pSB1C3, via RE cloning.</span></p><br />
<p style="text-align: center;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 20px; font-weight: bold; line-height: 19.5px; text-align: start;">Lpp-OmpA-Linker (pSB1C3)</span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">Outer membrane protein A (OmpA) is a common outer membrane protein found in bacteria that serves various purposes. For our purposes, the beta-barrel transmembrane domain can be utilized to anchor otherwise soluble proteins in the outer cell membrane. Lpp serves to direct the fusion protein, while a unstructured linker (GGGSGGGS) serves to separate OmpA from its cargo in order to avoid interactions between the two proteins.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">The expressed protein is structurally identical to BBa_K103006. This version of the biobrick has been moved into the new backbone, pSB1C3, which contains the gene for chloramphenicol resistance. This resistance gene contains an internal SacI site, making the SacI site at the end of the gene almost unusable for RE cloning. BBa_K1489003 mutagenizes this SacI site into a KasI in order to avoid this issue.</p><br />
<p><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;"> </span></p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="line-height: 12.0000009536743px; color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; text-decoration: underline;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">Part:BBa_K1489000 | </span></span><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap; text-decoration: underline;">pBAD-RBS-Bovine Galectin 1-B0012 Terminator</span></strong></span></p><br />
<p><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/parts/4/42/PSB1C3_-_BBa_K1489000.png" border="0" width="515" height="335" style="border: 0px; display: block; margin-left: auto; margin-right: auto;" /></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: 13px; line-height: 17.9400005340576px; white-space: pre-wrap;">A soluble his-tagged galectin from Bos taurus, expressed under Arabinose induction</span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px; text-align: center;"><span style="font-size: 20px; font-weight: bold;">pBAD-RBS-Bovine Galectin-1-B0012 Terminator</span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">Soluble beta-galactoside binding protein (galectin) originally from Bos taurus. Believed to expressed both intracellularly and extracellularly, where they serve to binding various types of beta-galactosides and initialize signal cascades. Believed to be a dimer in its active form, as is true with most mammalian galectins.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">This biobrick is under the pBAD promoter (BBa_K206000) with attached RBS (BBa_B0034), and utilizes an additional RNA polymerase terminator (BBa_B0012).</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">UMaryland 2014 is interested in utilizing this part to investigate whether E. coli can recognize and bind to surface carbohydrates on a marine pathogen. Other uses of this part lie in the recognition of various carbohydrate ligands and potential activation of signal transduction pathways.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;"> </p><br />
<p style="text-align: center;"> </p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="text-align: left; line-height: 17.9400005340576px; white-space: pre-wrap;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; text-decoration: underline;">Part:BBa_K1489004 | </span></span><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; line-height: 17.9400005340576px; white-space: pre-wrap; text-decoration: underline;">pBAD-RBS-Lpp-OmpA-Bovine Galectin 1-B0012 Terminator</span></strong></span></p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small; line-height: 17.9400005340576px; white-space: pre-wrap; text-decoration: underline;"><img src="https://static.igem.org/mediawiki/parts/0/0f/PSB1C3_-_BBa_K1489004.png" border="0" width="515" height="335" /></span></strong></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">His-tagged bovine galectin 1 attached to the c-terminus of OmpA, expressed under arabinose induction</span></span></p><br />
<p> </p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 20px; font-weight: bold; line-height: 19.5px; text-align: start;">pBAD-RBS-OmpA-Bovine Galectin-1-B0012 Terminator</span></span></p><br />
<p style="text-align: left;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 13px; line-height: 19.5px; text-align: start;">A fusion protein of OmpA (BBa_K1489002) fused to Bovine Galectin-1 (BBa_K1489001). Used to bind sugars at the outer cell membrane. Under pBAD regulation.</span></span></p><br />
<p style="text-align: left;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"> </span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"><span style="text-decoration: underline;"><strong>Part:BBa_K1489003 | Revised Lpp-OmpA</strong></span></span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"><span style="text-decoration: underline;"><strong><img src="https://static.igem.org/mediawiki/parts/9/90/PSB1C3_-_BBa_K1489003.png" border="0" width="515" height="335" /></strong></span></span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"> Biobrick 1, with a SacI site at the C-terminus replaced by a KasI site. This improvement was made in order to re-enable RE cloning; a SacI site exists in the chloramphenicol resistance gene.</span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 20px; font-weight: bold; line-height: 19.5px; text-align: start;">Lpp-OmpA-Linker with KasI site</span></span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">Outer membrane protein A (OmpA) is a common outer membrane protein found in bacteria that serves various purposes. For our purposes, the beta-barrel transmembrane domain can be utilized to anchor otherwise soluble proteins in the outer cell membrane. Lpp serves to direct the fusion protein, while a unstructured linker (GGGSGGGS) serves to separate OmpA from its cargo in order to avoid interactions between the two proteins.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">The expressed protein is almost structurally identical to BBa_K103006, with the terminal serine on the linker replaced by an alanine. This change was made in order to convert the previous SacI site (GAGCTC), which was unusable for RE cloning due to a second SacI site being present in the plasmid backbone, into a KasI site (GGCGCC). The KasI site codes for Gly-Ala, while the SacI site codes for Gly-Ser. This amino acid substitution is unlikely to impact the function of Lpp-OmpA-Linker in a major way.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;"> </p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;"> </p><br />
<p style="text-align: center;"><span style="text-decoration: underline;"><strong><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">Part:BBa_K1489001 | Bovine Galectin-1</span></span></strong></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/parts/4/4c/PSB1C3_-_BBa_K1489001.png" border="0" width="515" height="335" style="border: 0;" /></span></span></p><br />
<p style="text-align: center;"><span style="color: #141823; font-family: Helvetica, Arial, 'lucida grande', tahoma, verdana, arial, sans-serif; font-size: small;"><span style="line-height: 17.9400005340576px; white-space: pre-wrap;">This is the Bovine Galectin which should interact with beta-galactosides and intialize a signal cascase</span></span></p><br />
<p style="text-align: center;"><span style="color: #282828; font-family: arial, sans-serif; font-size: 20px; font-weight: bold; line-height: 19.5px; text-align: start;">Bovine Galectin-1</span></p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">Soluble beta-galactoside binding protein (galectin) originally from Bos taurus. Believed to expressed both intracellularly and extracellularly, where they serve to binding various types of beta-galactosides and initialize signal cascades. Believed to be a dimer in its active form, as is true with most mammalian galectins.</p><br />
<p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;">UMaryland 2014 is interested in utilizing this part to investigate whether E. coli can recognize and bind to surface carbohydrates on a marine pathogen. Other uses of this part lie in the recognition of various carbohydrate ligands and potential activation of signal transduction pathways.</p> <br />
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Nbb826
http://2014.igem.org/Team:UMaryland/project/background
Team:UMaryland/project/background
2014-10-16T21:01:08Z
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Background-Motivation <br />
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<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: center;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"><em><span style="text-decoration: underline;"><strong>Background</strong></span></em><br /></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">We at UMaryland iGEM are developing a biosensor to detect for the presence of an oyster pathogen, </span><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">Perkinsus marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">. This parasite is commonly found over the eastern shore of North America, including in Maryland’s own Chesapeake Bay. </span><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">, a unicellular protist, causes an oyster infection known as Dermo (Espinosa, Winnicki, &amp; Allam, 2013). The pathogen has been shown to infest oysters by entering their hemocytes via induced phagocytosis. When inside the hemocyte, the parasite is capable of resisting the reactive oxygen species generated by the oyster immune system and modulating the apoptosis of infected hemocyte cells to spread itself further (Espinosa et al., 2013). This leads to proliferation inside the oyster hemocyte, eventually lysing the host cell and releasing more </span><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"> (Allam et al., 2013). This process leads to massive tissue damage and, ultimately, the death of the oyster.</span></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">It is believed that </span><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">P. marinus </span><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">enters utilizing a galectin, a galactose-binding protein, on the hemocyte cell surface, CvGal1, which allows it to bind preferentially to </span><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"> (Schott, Pecher, Okafor, &amp; Vasta, 2003). Our aim is to exploit this property by expressing the galectin on the surface of </span><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">E. coli</span><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"> bacterial cells by attaching it to the C-terminus of an OmpA transmembrane protein. These bacteria will thus bind to any </span><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"> present in a sample of water. Our long-term goal is to incorporate these biosensors into a lab-on-a-chip device, requiring only small samples and allowing quick response times for detecting this pathogen.</span></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">This year, we have focused on working with the bovine galectin-1 in place of CvGal1. At only 14 kDa in size, the bovine galectin was chosen as a simpler candidate for expression on the surface of the membrane (Liao, Kapadia, Ahmed, Vasta, &amp; Herzberg, 1994). Furthermore, this bovine galectin-1 was one of three galectins used for the homology modeling of CvGal1 due to their similar binding domains (Tasumi &amp; Vasta, 2007). Though bovine galectin-1 is a two-domain galectin, compared to the four domains of CvGal1 (Feng et al., 2013), it is believed that the bovine galectin has potential to bind the same ligand present on </span><span style="font-size: 15px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"> that CvGal1 binds. Additionally, we have optimized the bovine galectin-1 gene, most notably for better protein folding and BioBrick friendliness.</span></p><br />
<p style="line-height: 12.0000009536743px;"> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">Although laboratory methods for Dermo detection using PCR exist, they are currently not capable of detecting the disease in real time. In the time needed to conduct a laboratory test, Dermo will have easily spread to new locations. If we can pinpoint the location of Dermo on-site, then we can take action ultimately to eliminate the parasite from eastern oyster habitats.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">References</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">Allam, B., Carden, W. E., Ward, J. E., Ralph, G., Winnicki, S., &amp; Espinosa, E. P. (2013). Early host-pathogen interactions in marine bivalves: Evidence that the alveolate parasite Perkinsus marinus infects through the oyster mantle during rejection of pseudofeces. </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">Journal of Invertebrate Pathology</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">, </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">113</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">(1), 26-34.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">Espinosa, E. P., Winnicki, S., &amp; Allam, B. (2013). Early host–pathogen interactions in a marine bivalve: Crassostrea virginica pallial mucus modulates Perkinsus marinus growth and virulence. </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">Diseases of Aquatic Organisms</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">, </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">104</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">(3), 237-247.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">Feng, C., Ghosh, A., Amin, M. N., Giomarelli, B., Shridhar, S., Banerjee, A., ... &amp; Vasta, G. R. (2013). The galectin CvGal1 from the eastern oyster (Crassostrea virginica) binds to blood group A oligosaccharides on the hemocyte surface. </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">Journal of Biological Chemistry</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">, </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">288</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">(34), 24394-24409.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">Liao, D. I., Kapadia, G., Ahmed, H., Vasta, G. R., &amp; Herzberg, O. (1994). Structure of S-lectin, a developmentally regulated vertebrate beta-galactoside-binding protein. </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">Proceedings of the National Academy of Sciences</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">, </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">91</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">(4), 1428-1432.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">Tasumi, S., &amp; Vasta, G. R. (2007). A galectin of unique domain organization from hemocytes of the Eastern oyster (Crassostrea virginica) is a receptor for the protistan parasite Perkinsus marinus. </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">The Journal of Immunology</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">, </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">179</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">(5), 3086-3098.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">Schott, E. J., Pecher, W. T., Okafor, F., &amp; Vasta, G. R. (2003). The protistan parasite Perkinsus marinus is resistant to selected reactive oxygen species. </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">Experimental Parasitology</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">, </span><span style="font-size: 11px; font-family: Arial; color: #000000; font-style: italic; vertical-align: baseline; white-space: pre-wrap;">105</span><span style="font-size: 11px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">(3-4), 232-240.</span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"> </p><br />
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Overview <br />
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<p style="margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: center;" dir="ltr"><em><span style="text-decoration: underline;"><strong><span style="vertical-align: baseline;"><span style="color: #000000; font-family: Arial;"><span style="font-size: 15.5555562973022px; line-height: 19.1666679382324px; white-space: pre-wrap;">Overview</span></span></span></strong></span></em></p><br />
<p style="margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: center;" dir="ltr"><em><span style="text-decoration: underline;"><strong><span style="vertical-align: baseline;"><span style="color: #000000; font-family: Arial;"><span style="font-size: 15.5555562973022px; line-height: 19.1666679382324px; white-space: pre-wrap;"><img src="https://static.igem.org/mediawiki/parts/f/f2/Umarylandigeminfographic.jpg" border="0" width="402" height="256" /></span></span></span></strong></span></em></p><br />
<p style="margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: left;" dir="ltr"><span style="vertical-align: baseline;"><span style="color: #000000; font-family: Arial;"><span style="font-size: 15.5555562973022px; line-height: 19.1666679382324px; white-space: pre-wrap;">The UMaryland iGem team opted for a project that hits close to home; with the Chesapeake Bay in our backyard, we have the potential to make a direct impact on the community. As Maryland’s most important natural landmark, the Chesapeake Bay is an integral part of our state’s identity. The bay is home to a diverse ecosystem, and is an economic boon to the area, supporting both the seafood industry and local tourism. Since the health of the bay is important, we looked into current problems in need of attention. In particular, the predicament of the Chesapeake’s oysters piqued our interest. As oyster populations declined over the years, the absence of this filtration system left a host of problems, including decreased water clarity and rising chemical pollutant levels. If the oyster populations can be maintained, it will improve the overall health of the Chesapeake. One way to combat oyster fatalities is to develop a way to monitor Dermo infestations. </span></span> </span></p><br />
<p style="text-align: left;"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"> </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: center;" dir="ltr"> </p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: center;" dir="ltr"><em><strong><span style="text-decoration: underline;"><span style="font-size: 15px; font-family: Arial; color: #000000; font-variant: normal; text-decoration: underline; vertical-align: baseline; white-space: pre-wrap; background-color: #ffffff;">Applications in the lab</span></span></strong></em></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">It is believed that </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">P. marinus </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">enters oyster hemocytes utilizing a galectin on the cell’s surface, CvGal1, that allows binds preferentially to </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">. Our aim is to exploit CvGal1’s ability to bind to </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">P. marinus</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> in order to design a biosensor for the parasite. By expressing the galectin on the surface of </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">E. coli</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> and triggering a signal transduction cascade within the bacterial cell, the </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">E. coli</span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> could produce Green Fluorescent Protein. Thus, </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">P. marinus </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">binding to CvGal1 would trigger the expression of GFP within </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: italic; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">E. coli, </span><span style="font-size: 15px; font-family: Arial; color: #000000; background-color: #ffffff; font-weight: normal; font-style: normal; font-variant: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;"> creating a visual marker for the parasite. The majority of this project will be accomplished through genetic cloning and bacterial transformation. Although laboratory methods for Dermo detection using Polymerase Chain Reaction exist, they are currently not capable of detecting the disease in real time. In the time needed to conduct a laboratory test, Dermo could have easily spread to new locations. This biosensor would help to accurately pinpoint Dermo infestation levels in real time. If we can pinpoint the location of Dermo, as elucidated in this research proposal, then we can ultimately develop a biosensor that can react to Dermo with targeted destruction. </span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;"><strong id="docs-internal-guid-809a97bc-f5c8-6033-4262-354a17c98c1d" style="font-weight: normal;"><br /><br /></strong></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: center;" dir="ltr"><span style="text-decoration: underline;"><strong><em><span style="font-size: 15px; font-family: Arial; color: #000000; font-variant: normal; text-decoration: underline; vertical-align: baseline; white-space: pre-wrap; background-color: transparent;">Brief overview of servicing the community</span></em></strong></span></p><br />
<p style="line-height: 1.15; margin-top: 0pt; margin-bottom: 0pt; margin-right: 4pt; text-align: justify;" dir="ltr"><span style="font-size: 15px; font-family: Arial; color: #000000; vertical-align: baseline; white-space: pre-wrap;">We having been traveling around Maryland conducting various community outreach efforts by visiting local schools and oyster farms. During each visit, members present our project and spread awareness on the current state of the Chesapeake Bay, as well as ways individuals can help service the Bay. Some presentations are made on synthetic biology to increase student participation and interest in the sciences. We also have a committee dedicated to creating an educational computer game that teaches others about the issues that the Chesapeake Bay currently faces. It is our team’s hope that our research results to be presented at the iGEM jamboree will have a profound and positive impact on the health of the Chesapeake Bay and oyster populations worldwide</span></p><br />
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http://2014.igem.org/Team:UMaryland/team/sponsors
Team:UMaryland/team/sponsors
2014-10-16T20:59:22Z
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href="https://2014.igem.org/Team:UMaryland/project/futureapplications"><span class="yjm_has_none"><span class="yjm_title">future applications</span></span></a></span></li><li class="item301 level1 lilast"><span class="mymarg"><a class="yjanchor last" href="https://igem.org/Safety/Safety_Form?team_id=1489"><span class="yjm_has_none"><span class="yjm_title">Safety</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li><li class="haschild item234 level0"><span class="child"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/notebook"><span class="yjm_has_none"><span class="yjm_title">Notebook</span></span></a></span><ul class="subul_main level1"><li class="bl"></li><li class="tl"></li><li class="tr"></li><li class="item303 level1 first lilast"><span class="mymarg"><a class="yjanchor firstlast" href="https://2014.igem.org/Team:UMaryland/notebook"><span class="yjm_has_none"><span class="yjm_title">Wetlab Notebook</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li><li class="haschild item123 level0"><span class="child"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/humanprac/overview"><span class="yjm_has_none"><span class="yjm_title">Human Practices</span></span></a></span><ul class="subul_main level1"><li class="bl"></li><li class="tl"></li><li class="tr"></li><li class="item106 level1 first"><span class="mymarg"><a class="yjanchor first" href="https://2014.igem.org/Team:UMaryland/humanprac/overview"><span class="yjm_has_none"><span class="yjm_title">Overview | Mission Statement</span></span></a></span></li><li class="haschild item129 level1"><span class="child subparent"><a class="yjanchor grouptitle" href="https://2014.igem.org/Team:UMaryland/humanprac/publicschool"><span class="yjm_has_none"><span class="yjm_title">Public School Outreach</span></span></a></span><ul class="subul_main level2"><li class="bl"></li><li class="tl"></li><li class="tr"></li><li class="item134 level2 first lilast"><span class="mymarg"><a class="yjanchor firstlast" href="https://2014.igem.org/Team:UMaryland/humanprac/publicschool"><span class="yjm_has_none"><span class="yjm_title">HighSchool</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li><li class="item131 level1"><span class="mymarg"><a class="yjanchor " href="https://2014.igem.org/Team:UMaryland/humanprac/community"><span class="yjm_has_none"><span class="yjm_title">Community Interaction</span></span></a></span></li><li class="item132 level1 lilast"><span class="mymarg"><a class="yjanchor last" href="https://2014.igem.org/Team:UMaryland/humanprac/publicawareness"><span class="yjm_has_none"><span class="yjm_title">Public Awareness</span></span></a></span></li><li class="right"></li><li class="br"></li></ul></li></ul></div><br />
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Sponsors <br />
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<p>We would like to thank all of our wonderful sponsors, without which our work would not be possible!</p><br />
<p><img src="https://static.igem.org/mediawiki/parts/c/c4/UMIDT.png" border="0" width="372" height="97" style="border: 0;" /></p><br />
<p><img src="https://static.igem.org/mediawiki/parts/5/5c/Qiagen.png" border="0" width="153" height="129" style="border: 0;" /><img src="https://static.igem.org/mediawiki/parts/5/55/UMNEB.jpg" border="0" width="298" height="116" style="border: 0;" /><img src="https://static.igem.org/mediawiki/parts/b/b0/UMClark.jpg" border="0" width="287" height="131" style="border: 0;" /><img src="https://static.igem.org/mediawiki/parts/c/c1/UMNationalScholarshipsOffice.jpg" border="0" alt="National Scholarships Office" width="525" height="70" style="border: 0;" /></p><br />
<p><img src="https://static.igem.org/mediawiki/parts/4/4f/UMILS.png" border="0" alt="Home" width="441" height="128" style="border: 0;" /> <img src="https://static.igem.org/mediawiki/parts/b/b1/UMCMNS.png" border="0" width="577" height="79" style="border: 0;" /> <p><img src="https://static.igem.org/mediawiki/parts/f/fb/ILS_text.jpg" border="0" alt="Home" width="250" height="100" style="border: 0;" /> </p> <br />
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<span class="yjmm_select" id="yjmm_selectid">Sponsors</span><br />
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<option value="/iGemWiki/">&nbsp;Homepage</option><br />
<option value="https://2014.igem.org/Team:UMaryland/photos">&nbsp;Photos</option><br />
<option value="https://2014.igem.org/Team:UMaryland/meetourteam">&nbsp;Team</option><br />
<option value="/iGemWiki/index.php/teaminfo/meet-the-team-members">&nbsp;--Meet Our Team Members</option><br />
<option value="https://igem.org/Team.cgi?id=1489">&nbsp;--iGEM Page</option><br />
<option value="http://localhost/iGemWiki/index.php/teaminfo/attributions">&nbsp;--Attributions</option><br />
<option value="https://2014.igem.org/Team:UMaryland/team/sponsors" selected="selected">&nbsp;--Sponsors</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/overview">&nbsp;Project</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/overview">&nbsp;--Overview</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/background">&nbsp;--Background</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/biobricks">&nbsp;--parts (BioBricks)</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/results">&nbsp;--Results</option><br />
<option value="https://2014.igem.org/Team:UMaryland/project/futureapplications">&nbsp;--future applications</option><br />
<option value="https://igem.org/Safety/Safety_Form?team_id=1489">&nbsp;--Safety</option><br />
<option value="https://2014.igem.org/Team:UMaryland/notebook">&nbsp;Notebook</option><br />
<option value="https://2014.igem.org/Team:UMaryland/notebook">&nbsp;--Wetlab Notebook</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/overview">&nbsp;Human Practices</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/overview">&nbsp;--Overview | Mission Statement</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicschool">&nbsp;--Public School Outreach</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicschool">&nbsp;---HighSchool</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/community">&nbsp;--Community Interaction</option><br />
<option value="https://2014.igem.org/Team:UMaryland/humanprac/publicawareness">&nbsp;--Public Awareness</option><br />
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