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Team:UMaryland/project/biobricks
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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 id="current" class=" active item144 level1"><span class="mymarg"><a class="yjanchor activepath " 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 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> |
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<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> | <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> | ||
<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> | <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> | ||
| - | <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. | + | <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> |
<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> | <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> | ||
<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> | <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> | ||
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<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> | <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> | ||
<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> | <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> | ||
| - | <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. | + | <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> |
| - | <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> | + | <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> |
| - | + | <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> | |
| - | + | <p style="margin: 0.4em 0px 0.5em; line-height: 19.5px; color: #282828; font-family: arial, sans-serif; font-size: 13px;"> </p> | |
| - | + | <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> | |
| - | + | <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> | |
| - | + | <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> | |
| - | + | <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> | |
| - | + | <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> | |
| - | + | <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> | |
| - | + | <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> | |
| - | + | <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> | |
| + | <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> | ||
| + | <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> | ||
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Latest revision as of 20:40, 17 October 2014
Biobricks
Part:BBa_K1489002 | Lpp-OmpA in new backbone
We moved an existing biobrick, BBa_K103006, from an old plasmid backbone, pSB1A2, into the current backbone, pSB1C3, via RE cloning.
Lpp-OmpA-Linker (pSB1C3)
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.
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.
Part:BBa_K1489000 | pBAD-RBS-Bovine Galectin 1-B0012 Terminator
A soluble his-tagged galectin from Bos taurus, expressed under Arabinose induction
pBAD-RBS-Bovine Galectin-1-B0012 Terminator
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.
This biobrick is under the pBAD promoter (BBa_K206000) with attached RBS (BBa_B0034), and utilizes an additional RNA polymerase terminator (BBa_B0012).
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.
Part:BBa_K1489004 | pBAD-RBS-Lpp-OmpA-Bovine Galectin 1-B0012 Terminator
His-tagged bovine galectin 1 attached to the c-terminus of OmpA, expressed under arabinose induction
pBAD-RBS-OmpA-Bovine Galectin-1-B0012 Terminator
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.
Part:BBa_K1489003 | Revised Lpp-OmpA
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.
Lpp-OmpA-Linker with KasI site
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.
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.
Part:BBa_K1489001 | Bovine Galectin-1
This is the Bovine Galectin which should interact with beta-galactosides and intialize a signal cascase
Bovine Galectin-1
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.
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.
Part:BBa_K1489005: Crasseoterra virginica Galectin 1 (CvGal1)
Crasseoterra virginica Galectin 1 (CvGal1)
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.
Part:BBa_K1489006: OmpA-CvGal1
OmpA-CvGal1
CvGal1 fused to OmpA in order to anchor it in the membrane.
About Umaryland
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!
