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http://2014.igem.org/Team:Stony_Brook/Results
Team:Stony Brook/Results
2014-10-18T03:51:05Z
<p>Hliu1423: </p>
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<div id="yellow_header"> <a href="http://www.stonybrook.edu/">Stony Brook University</a></div><br />
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<div id="blue_header"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/1/1e/Stony_Brook_Apis-biotics_header.png"/ id="apis-biotics"/></a><br />
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<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/7/78/Stony_Brook_NavIcon2_Home.png"/></a><br />
<p> Home </p><br />
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<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook/Team"><img src="https://static.igem.org/mediawiki/2014/6/66/Stony_Brook_NavIcon2_Team.png"/></a><br />
<p> Team </p><br />
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<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook/Project"><img src="https://static.igem.org/mediawiki/2014/9/99/Stony_Brook_NavIcon2_Project.png"/></a><br />
<p> Project </p><br />
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<p> Notebook </p><br />
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<a href="https://2014.igem.org/Team:Stony_Brook/Results"> <img src="https://static.igem.org/mediawiki/2014/0/04/Stony_Brook_NavIcon2_Results.png"/></a><br />
<p>Results</p><br />
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<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook/Outreach"><img src="https://static.igem.org/mediawiki/2014/8/80/Stony_Brook_NavIcon2_Outreach.png"/></a><br />
<p> Outreach </p><br />
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<div id="yellow_navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Safety"> <img src="https://static.igem.org/mediawiki/2014/0/07/Stony_Brook_NavIcon2_Safety.png"/></a><br />
<p>Safety</p><br />
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<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook/Attributions"><img src="https://static.igem.org/mediawiki/2014/0/03/Stony_Brook_NavIcon2_Acknowledgements.png"/></a><br />
<p> Attributions </p><br />
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<div id="project"><p>Results</p> <br />
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<h4> HIGHLIGHTS </h4><br />
<p><ul><li>We successfully produced melittin in our <i>E. coli</i> from honeybee DNA!</li><br />
<li> We were able to build and test our biosensor with our mCherry fluorescence marker!</li><br />
<li> We came up with some mathematical models to describe our circuit!</li><br />
<li> Check out our Biobricks <a href="http://parts.igem.org/Part:BBa_K1382000"> here</a> and <a href="http://parts.igem.org/Part:BBa_K1382001"> here</a>!</li></ul></p></div><br />
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<label for="tab-1" class="tab-label-1">GST-Melittin</label><br />
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<label for="tab-2" class="tab-label-2">RhlR Biosensor</label><br />
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<label for="tab-3" class="tab-label-3">Mathematical Modeling</label> <br />
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<h4>THE BUILDING PROCESS</h4><br />
<p>We used a GST plasmid (GST-C-His +) so that melittin could remain in its inactive form when attached to the GST tag. A TEV cleavage site was included in between the GST protein and the melittin peptide sequence so that upon exogenous addition of TEV protease, melittin will be cleaved from GST, thus activating the protein.<figure class="left"><img src="https://static.igem.org/mediawiki/2014/f/fa/Stony_Brook_Results_Dead_bees.jpg" /><figcaption> Our bees being dissected! </figcaption></figure></p><p> In order to find the gene within the originating organism, we obtained samples of Apis mellifera, amd performed an RNA extraction of the venom gland of the honeybee. After using reverse transcriptase to get the cDNA library, we used PCR to search the cDNA library for prepromelittin precursor to melittin. </p> <br />
<figure class="right"><img src="https://static.igem.org/mediawiki/2014/4/4b/Stony_Brook_Results_Tev_melittin.jpg" /><figcaption> PCR Screening for TEV-Melittin for a band around 109 base pairs </figcaption></figure><br />
<p>After multiple cDNA searches, we found the correct sized band for prepromelittin for 4 out of 6 of our samples. Once we saw we had the correct ~253 band of prepromelittin, we performed a gel extraction and used the DNA for another PCR.</p><br />
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<p>We used this second PCR to add the TEV cleavage site sequence and amplify out the melittin portion, excluding the pre- and pro- precursor sequences. Once we saw the correct band of ~100, we were ready to ligate our insert into the digested GST plasmid. </p> <br />
<p>Next, we wanted to induce our system with IPTG lac promoter to see if we could produce melittin, and so we grew out a culture, induced it, and performed SDS page alongside just the original GST plasmid to see if a size of ~30kDa would show up on the gel, which it did!</p><br/><br/><br />
<figure class="center"><img src="https://static.igem.org/mediawiki/2014/6/63/Stony_Brook_Results_Prepromelittin_and_tev_melittin.png" /><figcaption>Prepromelittin amplified out on two gels on the left, and TEV-Melittin on the right</figcaption></figure> <br />
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<figure class="center"><img src="https://static.igem.org/mediawiki/2014/1/1a/Stony_Brook_Results_Gst_melittin_sds_page.png" /><figcaption> SDS page showing a band for GST-melittin at the red arrow in the right four lanes, with a GST plasmid control in the third lane from the left</figcaption></figure><br/><br/><br />
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<h4> TESTING THE BIOACTIVITY OF MELITTIN </h4><br />
<figure> <img src="https://static.igem.org/mediawiki/2014/a/a9/Stony_Brook_Results_Melittin_plates.png" /></figure><br />
<figure> <img src="https://static.igem.org/mediawiki/2014/4/44/Stony_Brook_Results_Melittin_plates_after.png" /><figcaption> <i>E. coli </i> before and after the addition of melittin solution in an "x" shape.</figcaption></figure><br />
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<h4> TESTING THE RESPONSE OF OUR RHLR BIOSENSOR </h4><br />
<figure class="center"><img src="https://static.igem.org/mediawiki/2014/1/1b/Stony_Brook_Results_Gfp_fluorescene_activated_by_c4hsl.png" /></figure><br/><br/><p>The initial testing of the 2011 Northwestern construct. 50uM of C4-HSL was added along with a negative control and GFP fluorescence was tracked over a period of seven hours. The A and B refer to two separate colonies that were picked off the plate. Everything worked as expected during log phase. However, once the negative control began approaching saturation, it began expressing GFP indicating that it may actually be a leaky promoter. This is useful to know so that once our system is complete, we know that we need to have the cells in log or lag phase to avoid false positives. </p><br/><br/><br />
<figure class="center"><img src="https://static.igem.org/mediawiki/2014/c/c4/Stony_Brook_Results_Zero_um_c4hsl.png" /></figure><p>The data here is showing that GFP is being expressed after approximately 120 to 140 minutes in our initial test when no amount of C4-HSL was added</p><br/><br/><br />
<figure class="center"><img src="https://static.igem.org/mediawiki/2014/3/31/Stony_Brook_Results_Growth_curve_initial_gfp_test.png" /></figure><p>This growth curve is showing that fluorescence is being detected in the presence of C4-HSL when the cells are in log phase. However, it is also showing that the cells which did not receive the C4-HSL treatment are expressing GFP when they reach saturation.</p><br/><br/><br />
<figure class="center"><img src="https://static.igem.org/mediawiki/2014/d/d4/Stony_Brook_Results_Cotransfected_e_coli.png" /></figure><br/><br/><p>This graph is showing that our cells are responding to the C4-HSL treatment. These cells were cotransfected with two plasmids. One plasmid has the Rhl promoter with mCherry acting as a reporter. The otherplasmid has our RhlR protein generator constitutively expressed.</p><br/><br/><br />
<figure class="center"><img src="https://static.igem.org/mediawiki/2014/4/40/Stony_Brook_Results_Growth_curve_negative_control.png"</figure><br/><br/><p>The Northwestern was tested once again and we still saw the same results: GFP was expressed once cells reached saturation. Despite the low level of expression, it can still lead to false positives in the future.</p><br/><br/><br />
<figure class="center"><img src="https://static.igem.org/mediawiki/2014/7/74/Stony_Brook_Results_Rhl_promoter.png" /><br/><br/></figure><p>To test to see if the Rhl promoter is in fact a leaky promoter, cells were transfected with plasmids containing just the Rhl promoter and mCherry. The results show that once cells reach saturation, fluorescence is detected, suggesting that Rhl is in fact a leaky promoter.</p><br />
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<h4> MATHEMATICAL MODELING </h4><br />
<figure class="center"><img src="https://static.igem.org/mediawiki/2014/6/6b/Stony_Brook_Math_Model_Network_model.png" /></figure><br />
<br/><p> Legend:</p><br />
<p>Ae= Extracellular AHL </p><br />
<p>Ai= Intracellular AHL</p><br />
<p>d = AHL diffusion rate</p><br />
<p>g = decay rate of intracellular AHL</p><br />
<p>R= RhlR </p><br />
<p>r = RhlR synthesis rate</p><br />
<p>? = RhlR decay rate</p><br />
<p>C= AHL: RhlR complex (an active transcription factor)</p><br />
<p>b = binding rate of AHL with RhlR </p><br />
<p>� = dissociation rate of AHL:RhlR complex</p><br />
<p>G = Melittin DNA</p><br />
<p>Cg= AHL + RhlR + Melittin DNA complex</p><br />
<p>Kon = binding rate of the AHL:RhlR complex to the Melittin DNA</p><br />
<p>Koff = unbinding rate of AHL:RhlR complex from the Melittin DNA</p><br />
<p>M = Melittin mRNA</p><br />
<p>t = transcription rate for melittin</p><br />
<p>m = Melittin mRNA decay rate</p><br />
<p>P = Melittin Protein</p><br />
<p>T = translation rate for melittin</p><br />
<p>z = Protein Melittin�s decay rate</p><br />
<figure class="center"><img src="../../wiki pictures/results/modeling/parameters.png" /></figure><br />
<figure class="center"><img src="../../wiki pictures/results/modeling/Picture1.png" /></figure><br />
<p>Pseudomonas aeruginosa produces AHLe (external C4HSL) and we assumed it is at a constant rate. Only part of AHLe diffuses into the membrane AHLi (internal C4HSL). AHL has a higher concentration out of the cell than inside. Due to the chemical driving force, there is a fast increase in diffusion of AHL into the cell. AHL eventually reaches chemical equilibrium and the AHLi remains relatively constant. The cell produces RhlR, but RhlR binds with AHL to form an AHLi-RHL complex, hence the amount of �free� RhlR decreases in the cell as it is forming the complex. The AHLi-RHL complex activates the transcription of melittin, hence increases the presence of melittin mRNA (MELT_mRNA). The melittin mRNA barely decreases over time because the degradation rate is slow and melittin mRNA is converted into protein melittin without being broken down. Melittin protein is produced at a fast rate and eventually the rate of production becomes constant.</p><br />
<figure class="center"><img src="https://static.igem.org/mediawiki/2014/a/a4/Stony_Brook_Math_Model_Eq_1.png" /></figure><br />
<figure class="center"><img src="https://static.igem.org/mediawiki/2014/d/da/Stony_Brook_Math_Model_Eq_2_3.png" /></figure><br />
<figure class="center"><img src="https://static.igem.org/mediawiki/2014/e/e9/Stony_Brook_Math_Model_Eq_4.png" /></figure><br />
<figure class="center"><img src="https://static.igem.org/mediawiki/2014/e/e3/Stony_Brook_Math_Model_Eq_5_6.png" /></figure><br />
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padding: 0;<br />
}<br />
<br />
body .firstHeading {<br />
margin: 0;<br />
}<br />
<br />
div#contentSub {<br />
color: inherit;<br />
font-size: inherit;<br />
line-height: inherit;<br />
margin: 0;<br />
width: auto;<br />
}<br />
<br />
div#catlinks {<br />
background-color: transparent;<br />
border: 0;<br />
clear: both;<br />
margin: 0;<br />
padding: 0;<br />
}<br />
<br />
<br />
/* Start footer reset =================================================== */<br />
<br />
div#footer-box {<br />
background-color: transparent;<br />
border: 0;<br />
margin: 0;<br />
padding: 0;<br />
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}<br />
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div#footer {<br />
color: inherit;<br />
font-size: inherit;<br />
text-align: left;<br />
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div#f-poweredbyico, div#f-copyrightico {<br />
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div#footer li {<br />
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<br />
#contentSub, #footer-box, #catlinks, #search-controls, #p-logo, .printfooter, .firstHeading,.visualClear {display: none;}<br />
<br />
<br />
</style><br />
</html></div>
Hliu1423
http://2014.igem.org/Team:Stony_Brook
Team:Stony Brook
2014-10-18T03:44:41Z
<p>Hliu1423: </p>
<hr />
<div>{{CSS/Main}}<br />
<br />
<html xmlns="http://www.w3.org/1999/xhtml"><br />
<br />
<br />
<style type="text/css"><br />
<br />
#contentSub, #footer-box, #catlinks, #search-controls, #p-logo, .printfooter, .firstHeading,.visualClear {display: none;} <br />
<br />
body {<br />
background-image: url(https://static.igem.org/mediawiki/2014/a/a5/Stony_brook_home_background.png);<br />
background-repeat:no-repeat;<br />
background-position: center center;<br />
background-attachment: fixed;<br />
-webkit-background-size: cover;<br />
-moz-background-size: cover;<br />
-o-background-size: cover;<br />
background-size: cover;<br />
}<br />
#header {<br />
width: 672px;<br />
position: absolute;<br />
top:29%;<br />
left:50%;<br />
margin-left: -310px;<br />
margin-top:-50px;<br />
}<br />
<br />
#navigation {<br />
width: 90px;<br />
padding: 9px;<br />
float: left;<br />
color: #FFF;<br />
font-family:"Trebuchet MS", Arial, Helvetica, sans-serif;<br />
text-align: center;<br />
font-size:small;<br />
font-weight:bold;<br />
}<br />
<br />
#navigation p{<br />
display:none;<br />
text-align:center;<br />
}<br />
<br />
#navigation a:hover + p{<br />
display: block;<br />
}<br />
<br />
img:hover{<br />
opacity: 0.8;<br />
filter: alpha(opacity=80);<br />
}<br />
<br />
#iGEM_logo {<br />
position: absolute;<br />
left: 90%;<br />
top: 2%;<br />
}<br />
<br />
#navigation_container {<br />
width: 1000 px;<br />
position: absolute;<br />
top: 37%;<br />
right: 50%;<br />
margin-right: -450px;<br />
}<br />
<br />
<br />
<br />
a{<br />
outline: 0;<br />
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font-size: x-small;<br />
vertical-align: middle;<br />
font-weight: bold;<br />
color: #FFF;<br />
}<br />
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#login a:hover{<br />
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}<br />
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padding-bottom: 5px;<br />
}<br />
<br />
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bottom: 1%;<br />
left: 7%;<br />
padding-bottom: 5px;<br />
}<br />
<br />
#projectDescription{<br />
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bottom:25%;<br />
left:50%;<br />
width:584px;<br />
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margin-left:-265.6px;<br />
margin-bottom:-107px;<br />
overflow:auto;<br />
<br />
}<br />
#projectDescription h1, p{<br />
font-family:"Trebuchet MS", Arial, Helvetica, sans-serif;<br />
color:#FFFFFF;<br />
padding: 0px 20px 0px 20px;<br />
opacity:1;<br />
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#projectDescription h1{<br />
text-align:center;<br />
}<br />
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#projectDescription p{<br />
text-indent: 25px;<br />
}<br />
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</style><br />
<br />
</head><br />
<br />
<body><br />
<br />
<a href="https://igem.org/Main_Page"><img src="https://static.igem.org/mediawiki/2014/5/51/Stony_brook_igem_logo.png" alt= "iGEM logo" name="iGEM_logo" border="0px" id="iGEM_logo" /></a><br />
<br />
<a href="http://www.facebook.com/iGEMatstonybrook"><img src="https://static.igem.org/mediawiki/2014/0/0f/Stony_brook_fb_logo.png" alt= "facebook" name="facebook" border="0px" id="facebook" /></a></a><br />
<a href="http://twitter.com/iGEMSBU"><img src="https://static.igem.org/mediawiki/2014/b/b5/Stony_brook_twitter_logo.png" alt= "twitter" name="twitter" border="0px" id="twitter" /></a></a><br />
<br />
<br />
<div id="header"><a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/1/11/Stony_brook_header.png" alt="Stony Brook iGEM" border="0px"" /></a></div><br />
<br />
<div id="navigation_container"><br />
<br />
<div id="navigation"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/b/b8/Stony_Brook_NavIcon.Home.png" border="0px" /></a><br />
<p>Home</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Team"><img src="https://static.igem.org/mediawiki/2014/4/4a/Stony_Brook_NavIcon.Team.png" border="0px" /></a><br />
<p>Team</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Project"><img src="https://static.igem.org/mediawiki/2014/a/ab/Stony_Brook_NavIcon.Project.png" border="0px" /></a><br />
<p>Project</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Notebook"><img src="https://static.igem.org/mediawiki/2014/b/b5/Stony_Brook_NavIcon.Notebook.png" border="0px"/></a><br />
<p>Notebook</p><br />
</div><br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Results"> <img src="https://static.igem.org/mediawiki/2014/c/c3/Stony_Brook_NavIcon.Results.png"/></a><br />
<p>Results</p><br />
</div><br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Outreach"> <img src="https://static.igem.org/mediawiki/2014/e/e2/Stony_Brook_Navicon.Outreach.png" border="0px"/></a><br />
<p>Outreach</p><br />
</div> <br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Safety"> <img src="https://static.igem.org/mediawiki/2014/8/81/Stony_Brook_NavIcon.Safety.png"/></a><br />
<p>Safety</p><br />
</div> <br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Attributions"> <img src="https://static.igem.org/mediawiki/2014/5/5e/Stony_Brook_NavIcon.Acknowledgements.png" border="0px"/></a><br />
<p>Attributions</p><br />
</div> <br />
</div><br />
<br />
<br />
<!-- <div id="background"><br />
</div> --><br />
<br />
<!-- <div id="projectDescription"><br />
<h1> Project Description </h1><br />
<p> The threat of antibiotic resistance has been a concern since the discovery of penicillin. As antibiotics continue to be used to treat disease, pathogenic bacteria continue to develop resistance, eventually becoming irresponsive to multiple classes of antibiotics. This is a major problem for areas that must maintain sterile environments, such as hospitals or other health-care settings. Hospital-acquired infections (HAI) are already an existing issue that poses risks to people receiving or recovering from treatment. Gram-negative bacteria in particular are known to cause more HAIs as they are more resistant to antibiotics than gram-positive bacteria. Irresponsible use of antibiotics has only exacerbated this issue. Recent reports from the World Health Organization confirm that antibiotic resistance is no longer something to worry about in the future, but a current, global health threat. </p><br />
<br />
<p> Alternatives to modern antibiotics, such as antimicrobial peptides, are now being researched. Antimicrobial peptides, or AMPs, are peptides produced innately by the immune systems of certain organisms to fight off infection. Because they kill microorganisms through different mechanisms, they have the potential to be effective against antibiotic resistant bacteria. One such AMP is melittin, produced by honey bees and used in their venom. Melittin kills cells by forming pores in the cell membrane, eventually destabilizing the cell and causing cell lysis. Because melittin targets a conserved area of cells, bacteria cannot develop resistance to it as easily as different types of antibiotics. </p><br />
<br />
<p> Our project this summer is to transform E. coli with the melittin gene in honey bees, so that the E. coli may express the melittin peptide, thereby developing a means to mass-produce melittin as an alternative to today's antibiotics. We hope to optimize the production of melittin in E. coli, as well as to test the bioactivity of our produced melittin in comparison to its naturally-occurring counterpart. </p><br />
</div> --><br />
<br />
</body><br />
<br />
</html></div>
Hliu1423
http://2014.igem.org/Team:Stony_Brook
Team:Stony Brook
2014-10-18T03:43:56Z
<p>Hliu1423: </p>
<hr />
<div>{{CSS/Main}}<br />
<br />
<html xmlns="http://www.w3.org/1999/xhtml"><br />
<br />
<br />
<style type="text/css"><br />
<br />
#contentSub, #footer-box, #catlinks, #search-controls, #p-logo, .printfooter, .firstHeading,.visualClear {display: none;} <br />
<br />
body {<br />
background-image: url(https://static.igem.org/mediawiki/2014/a/a5/Stony_brook_home_background.png);<br />
background-repeat:no-repeat;<br />
background-position: center center;<br />
background-attachment: fixed;<br />
-webkit-background-size: cover;<br />
-moz-background-size: cover;<br />
-o-background-size: cover;<br />
background-size: cover;<br />
}<br />
#header {<br />
width: 672px;<br />
position: absolute;<br />
top:29%;<br />
left:50%;<br />
margin-left: -310px;<br />
margin-top:-50px;<br />
}<br />
<br />
#navigation {<br />
width: 90px;<br />
padding: 9px;<br />
float: left;<br />
color: #FFF;<br />
font-family:"Trebuchet MS", Arial, Helvetica, sans-serif;<br />
text-align: center;<br />
font-size:small;<br />
font-weight:bold;<br />
}<br />
<br />
#navigation p{<br />
display:none;<br />
text-align:center;<br />
}<br />
<br />
#navigation a:hover + p{<br />
display: block;<br />
}<br />
<br />
img:hover{<br />
opacity: 0.8;<br />
filter: alpha(opacity=80);<br />
}<br />
<br />
#iGEM_logo {<br />
position: absolute;<br />
left: 90%;<br />
top: 2%;<br />
}<br />
<br />
#navigation_container {<br />
width: 1000 px;<br />
position: absolute;<br />
top: 37%;<br />
right: 50%;<br />
margin-right: -450px;<br />
}<br />
<br />
<br />
<br />
a{<br />
outline: 0;<br />
}<br />
<br />
#login{<br />
width: 400px;<br />
height: 20px;<br />
background-color: #000;<br />
opacity: 0.7;<br />
filter: alpha(opacity=70);<br />
position: absolute;<br />
top: 0;<br />
left:0;<br />
}<br />
#login ul{<br />
list-style-type: none;<br />
margin: 0;<br />
padding: 0px;<br />
}<br />
<br />
#login li{<br />
float: left;<br />
text-align: center;<br />
}<br />
<br />
#login a{<br />
display: block;<br />
height: 20px;<br />
line-height: 20px;<br />
width: 100px;<br />
text-decoration: none;<br />
color: #FFF;<br />
font-family: "Trebuchet MS", Arial, Helvetica, sans-serif;<br />
font-size: x-small;<br />
vertical-align: middle;<br />
font-weight: bold;<br />
color: #FFF;<br />
}<br />
<br />
#login a:hover{<br />
color: #0F0;<br />
}<br />
<br />
.firstHeading {<br />
visibility: hidden;<br />
}<br />
<br />
#facebook{<br />
position: absolute;<br />
bottom: 1%;<br />
left: 1%;<br />
padding-bottom: 5px;<br />
}<br />
<br />
#twitter{<br />
position: absolute;<br />
bottom: 1%;<br />
left: 7%;<br />
padding-bottom: 5px;<br />
}<br />
<br />
#projectDescription{<br />
position:absolute;<br />
bottom:25%;<br />
left:50%;<br />
width:584px;<br />
height:320px;<br />
margin-left:-265.6px;<br />
margin-bottom:-107px;<br />
overflow:auto;<br />
<br />
}<br />
#projectDescription h1, p{<br />
font-family:"Trebuchet MS", Arial, Helvetica, sans-serif;<br />
color:#FFFFFF;<br />
padding: 0px 20px 0px 20px;<br />
opacity:1;<br />
}<br />
<br />
#projectDescription h1{<br />
text-align:center;<br />
}<br />
<br />
#projectDescription p{<br />
text-indent: 25px;<br />
}<br />
<br />
#background{<br />
position:absolute;<br />
bottom:25%;<br />
left:50%;<br />
width:584px;<br />
height:320px;<br />
background-color:#000;<br />
margin-left:-265.6px;<br />
margin-bottom:-107px;<br />
color: #FFF;<br />
opacity:0.6;<br />
filter:alpha(opacity=60);<br />
}<br />
<br />
</style><br />
<br />
</head><br />
<br />
<body><br />
<br />
<a href="https://igem.org/Main_Page"><img src="https://static.igem.org/mediawiki/2014/5/51/Stony_brook_igem_logo.png" alt= "iGEM logo" name="iGEM_logo" border="0px" id="iGEM_logo" /></a><br />
<br />
<a href="http://www.facebook.com/iGEMatstonybrook"><img src="https://static.igem.org/mediawiki/2014/0/0f/Stony_brook_fb_logo.png" alt= "facebook" name="facebook" border="0px" id="facebook" /></a></a><br />
<a href="http://twitter.com/iGEMSBU"><img src="https://static.igem.org/mediawiki/2014/b/b5/Stony_brook_twitter_logo.png" alt= "twitter" name="twitter" border="0px" id="twitter" /></a></a><br />
<br />
<br />
<div id="header"><a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/1/11/Stony_brook_header.png" alt="Stony Brook iGEM" border="0px"" /></a></div><br />
<br />
<div id="navigation_container"><br />
<br />
<div id="navigation"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/b/b8/Stony_Brook_NavIcon.Home.png" border="0px" /></a><br />
<p>Home</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Team"><img src="https://static.igem.org/mediawiki/2014/4/4a/Stony_Brook_NavIcon.Team.png" border="0px" /></a><br />
<p>Team</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Project"><img src="https://static.igem.org/mediawiki/2014/a/ab/Stony_Brook_NavIcon.Project.png" border="0px" /></a><br />
<p>Project</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Notebook"><img src="https://static.igem.org/mediawiki/2014/b/b5/Stony_Brook_NavIcon.Notebook.png" border="0px"/></a><br />
<p>Notebook</p><br />
</div><br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Results"> <img src="https://static.igem.org/mediawiki/2014/c/c3/Stony_Brook_NavIcon.Results.png"/></a><br />
<p>Results</p><br />
</div><br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Outreach"> <img src="https://static.igem.org/mediawiki/2014/e/e2/Stony_Brook_Navicon.Outreach.png" border="0px"/></a><br />
<p>Outreach</p><br />
</div> <br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Safety"> <img src="https://static.igem.org/mediawiki/2014/8/81/Stony_Brook_NavIcon.Safety.png"/></a><br />
<p>Safety</p><br />
</div> <br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Attributions"> <img src="https://static.igem.org/mediawiki/2014/5/5e/Stony_Brook_NavIcon.Acknowledgements.png" border="0px"/></a><br />
<p>Attributions</p><br />
</div> <br />
</div><br />
<br />
//**<div id="login"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook">page</a></li><br />
<li><a href="https://2014.igem.org/Talk:Team:Stony_Brook">discussion</a></li><br />
<li><a href="https://2014.igem.org/wiki/index.php?title=Team:Stony_Brook&amp;action=edit">view source</a></li><br />
<li><a href="https://2014.igem.org/wiki/index.php?title=Team:Stony_Brook&amp;action=history">history</a></li><br />
</ul><br />
</div>**//<br />
<br />
<!-- <div id="background"><br />
</div> --><br />
<br />
<!-- <div id="projectDescription"><br />
<h1> Project Description </h1><br />
<p> The threat of antibiotic resistance has been a concern since the discovery of penicillin. As antibiotics continue to be used to treat disease, pathogenic bacteria continue to develop resistance, eventually becoming irresponsive to multiple classes of antibiotics. This is a major problem for areas that must maintain sterile environments, such as hospitals or other health-care settings. Hospital-acquired infections (HAI) are already an existing issue that poses risks to people receiving or recovering from treatment. Gram-negative bacteria in particular are known to cause more HAIs as they are more resistant to antibiotics than gram-positive bacteria. Irresponsible use of antibiotics has only exacerbated this issue. Recent reports from the World Health Organization confirm that antibiotic resistance is no longer something to worry about in the future, but a current, global health threat. </p><br />
<br />
<p> Alternatives to modern antibiotics, such as antimicrobial peptides, are now being researched. Antimicrobial peptides, or AMPs, are peptides produced innately by the immune systems of certain organisms to fight off infection. Because they kill microorganisms through different mechanisms, they have the potential to be effective against antibiotic resistant bacteria. One such AMP is melittin, produced by honey bees and used in their venom. Melittin kills cells by forming pores in the cell membrane, eventually destabilizing the cell and causing cell lysis. Because melittin targets a conserved area of cells, bacteria cannot develop resistance to it as easily as different types of antibiotics. </p><br />
<br />
<p> Our project this summer is to transform E. coli with the melittin gene in honey bees, so that the E. coli may express the melittin peptide, thereby developing a means to mass-produce melittin as an alternative to today's antibiotics. We hope to optimize the production of melittin in E. coli, as well as to test the bioactivity of our produced melittin in comparison to its naturally-occurring counterpart. </p><br />
</div> --><br />
<br />
</body><br />
<br />
</html></div>
Hliu1423
http://2014.igem.org/Team:Stony_Brook/Project
Team:Stony Brook/Project
2014-10-18T01:26:30Z
<p>Hliu1423: </p>
<hr />
<div><!DOCTYPE HTML PUBLIC "-//W3C//DTD HTML 4.01 Transitional//EN" "http://www.w3.org/TR/html4/loose.dtd"><br />
<html xmlns="http://www.w3.org/1999/xhtml"><br />
<head><br />
<meta http-equiv="Content-Type" content="text/html; charset=utf-8" /><br />
<title>Untitled Document</title><br />
<style type="text/css"><br />
<br />
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<br />
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display:table;<br />
}<br />
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text-align:center;<br />
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background-color: #234678; <br />
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<body bgcolor="#ffffff"><br />
<div id="yellow_header"> <a href="http://www.stonybrook.edu/">Stony Brook University</a></div><br />
<br />
<div id="blue_header"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/1/1e/Stony_Brook_Apis-biotics_header.png"/ id="apis-biotics"/></a><br />
<br />
<div id="yellow_navigation_container"><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/7/78/Stony_Brook_NavIcon2_Home.png"/></a><br />
<p> Home </p><br />
</div><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook/Team"><img src="https://static.igem.org/mediawiki/2014/6/66/Stony_Brook_NavIcon2_Team.png"/></a><br />
<p> Team </p><br />
</div><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook/Project"><img src="https://static.igem.org/mediawiki/2014/9/99/Stony_Brook_NavIcon2_Project.png"/></a><br />
<p> Project </p><br />
</div><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook/Notebook"><img src="https://static.igem.org/mediawiki/2014/5/57/Stony_Brook_NavIcon2_Notebook.png"/></a><br />
<p> Notebook </p><br />
</div><br />
<div id="yellow_navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Results"> <img src="https://static.igem.org/mediawiki/2014/0/04/Stony_Brook_NavIcon2_Results.png"/></a><br />
<p>Results</p><br />
</div><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook/Outreach"><img src="https://static.igem.org/mediawiki/2014/8/80/Stony_Brook_NavIcon2_Outreach.png"/></a><br />
<p> Outreach </p><br />
</div><br />
<div id="yellow_navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Safety"> <img src="https://static.igem.org/mediawiki/2014/0/07/Stony_Brook_NavIcon2_Safety.png"/></a><br />
<p>Safety</p><br />
</div> <br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/0/03/Stony_Brook_NavIcon2_Acknowledgements.png"/></a><br />
<p> Attributions </p><br />
</div><br />
</div> <br />
</div><br />
<br />
<div id="project"><p>Project</p> <br />
</div><br />
<br />
<div id="abstract"> <h3>Abstract</h3><br />
<p>Pathogenic bacteria are becoming increasingly antibiotic-resistant due to misuse, overuse and abuse. In addition, infections caused by some harmful strains of bacteria, particularly gram-negative bacteria, cannot easily be treated with antibiotics or other common forms of treatment. Our intention is to engineer cells which can both produce an antimicrobial peptide and recognize the communication signals of such bacteria, using Pseudomonas aeruginosa as a model. Two plasmids, one which controls the production of our antimicrobial peptide melittin and the other which acts as the cell-signal receiver, will be inserted in nonpathogenic E. coli cells. This allows our E. coli to recognize the cells of Pseudomonas aeruginosa, and release a bacteria-killing compound in response. </p><br />
</div><br />
<div id="slideshow"><img src="https://static.igem.org/mediawiki/2014/a/ad/Stony_Brook_Project_Slideshow1.png" height="100%"></div><br />
<br />
<div id="contents"><br />
<section class="tabs"><br />
<input id="tab-1" type="radio" name="radio-set" class="tab-selector-1" checked="checked" /><br />
<label for="tab-1" class="tab-label-1">Antibiotic resistance</label><br />
<br />
<input id="tab-2" type="radio" name="radio-set" class="tab-selector-2" /><br />
<label for="tab-2" class="tab-label-2">P. aeruginosa</label><br />
<br />
<input id="tab-3" type="radio" name="radio-set" class="tab-selector-3" /><br />
<label for="tab-3" class="tab-label-3">Melittin</label><br />
<br />
<input id="tab-4" type="radio" name="radio-set" class="tab-selector-4" /><br />
<label for="tab-4" class="tab-label-4">Our Solution: Apis-Biotics</label><br />
<br />
<br />
<div class="clear-shadow"></div><br />
<br />
<div class="contents"><br />
<div class="content-1"><br />
<p><br />
<h4>WHEN ANTIBIOTICS FAIL</h4><br />
<br />
When Alexander Fleming was accepting his Nobel Prize in medicine for his discovery of penicillin, the world’s first antibiotic, he warned of a future where antibiotics are no longer effective. <sup><a href="#1">[1]</a></sup> The growth of antibacterial resistance—when bacteria no longer are affected by antibiotics—is quickly becoming a major public health concern. In 2014, the World Health Organization stated “A post-antibiotic era—in which common infections and minor injuries can kill—far from being an apocalyptic fantasy, is instead a very real possibility for the 21<sup>st</sup> century.” <sup><a href="#2">[2]</a></sup> </p><br />
<br />
<h4>THE MECHANISM OF ANTIBIOTIC RESISTANCE</h4><br />
<br />
<p>Antibiotic resistance is a natural phenomenon that occurs when mutations in bacterial replications render antibiotics ineffective. Many antibiotics kill bacteria by either preventing <br />
them from creating bacterial cell walls, affecting their DNA, RNA, or proteins, or changing their metabolism. However, through mutations, bacteria may develop the ability to break down <figure id="betalactamase"><br />
<img src="https://static.igem.org/mediawiki/2014/c/c9/Stony_Brook_Project_Beta_lactam_penicillin_action.png"><br />
<figcaption>Bacteria are constantly rebuilding their cell walls through peptidoglycan synthesis. Penicillin works by deactivating transpeptidase within the bacteria, preventing the cross-linking of peptidoglycan and weakening the cell wall, eventually causing lysis due to osmotic pressure. Bacteria can develop resistance to penicillin by producing β-lactamase, which breaks the beta-lactam ring in penicillin, deactivating it.</figcaption></figure>these antibiotics, prevent the antibiotic from entering the cell, or use efflux pumps in the cell membrane to transport the antibiotic back outside the cell. When antibiotics are used to kill off harmful bacteria, the resistant strain of bacteria remains and reproduces. With other strains <br />
gone, this resistant strain has less competition, allowing it to thrive.<br />
<br />
</p><br />
<br />
<p>However, what occurs more often than mutation-derived resistance is the spread of antibiotic resistance through horizontal gene transfer, which occurs when genetic material from bacteria, such as plasmids, transposons, or DNA are passed from cell to cell. Through this mechanism, bacteria can even develop multi-drug resistance. Even when a singular antibiotic is used for more than ten days, multidrug resistance develops to structurally unrelated drugs as the resistant bacteria recruit resistance genes from other bacteria in the environment, encouraging the development of “super-bugs.” In addition, some bacteria are intrinsically resistant to bacteria, such as gram-negative bacteria, which due to its thicker outer cell membrane are harder to treat with certain classes of antibiotics. In some cases, some strains of gram-negative bacteria are resistant to all antibiotics, even the <br />
newest families of antibiotics and antibiotics used as a last resort. <sup><a href="#3">[3]</a></sup><br />
<figure id="antibioticresistance"><img src="https://static.igem.org/mediawiki/2014/c/ce/Stony_Brook_Project_Antibiotic_resistance_selection.png"><figcaption>After an antibiotic is used, bacteria with resistance genes are selected while those without resistance die off, allowing resistant bacteria to better compete for resources and thrive</figcaption></figure><figure id="horizontal"><img src="https://static.igem.org/mediawiki/2014/4/43/Stony_Brook_Project_Horizontal_gene_transfer.png"><figcaption> Horizontal gene transfer: DNA for antibiotic resistance is transfered through conjugation.</figcaption></figure><br />
</p><br />
<br />
<br />
<p>The misuse of antibiotics in the healthcare environment contributes one avenue for the spread of antibiotic resistance. In one study done for the CDC, researchers found that almost 80% of hospitals misuse antibiotics, either giving inappropriate or redundant antibiotic combinations and intravenous antibiotics. <sup><a href="#4">[4]</a></sup> The use of antibiotics in food and agriculture also presents a significant problem. The Union of Concerned Scientists estimated in one report that 70% of all antibiotics used in the U.S. are used in the food and water of already-healthy livestock, in order to control disease. Many of these antibiotics don’t break down in the waste of livestock, leading to the growth of resistant bacteria which spreads as this waste is used as fertilizer, contaminating the soil and sources of water. This contamination can also be found coming from antibiotic manufacturing plants, as well as households which improperly dispose of unused or expired <br />
antibiotic pills, instead or returning them to a pharmacy. <sup><a href="#4">[5]</a></sup></p><br />
<br><br />
<br><br />
<br><br />
<br><br />
<br><br />
<br><br />
<br><br />
<br><br />
<br><br />
<br><br />
<br><br />
<br><br />
<br />
<h4>TO WHAT EXTENT?</h4><br />
<br />
<p>In its 2014 report on antibiotic resistance, the World Health Organization reported the current status of antibiotic resistance in seven major bacteria which cause disease in humans, including Escherichia coli, Klebsiella pneumonia, Staphylococcus aureus, and Streptococcus pneumoniae. These bacteria are common causes of infections within a hospital or community. If they become highly resistant to antibiotics, there would be significant repercussions for public health <br />
worldwide. </p><br />
<br />
<figure class="center"><img src='https://static.igem.org/mediawiki/2014/b/bc/Stony_Brook_Project_Antibiotic_resistance_chart.png'><figcaption> Data taken from WHO 2014 report on antimicrobial resistance </figcaption></figure><br />
<br />
<h4>WHY DOES IT MATTER?</h4><br />
<br />
<p>Antibiotic resistance is a global concern—it jeopardizes our ability to treat common infections, causing standard treatments to be replaced by more expensive drug therapies, making infectious diseases longer to treat and easier to spread, putting major surgeries, chemotherapy and organ transplants at a greater risk of being unsuccessful, and increasing death rate. <sup><a href="#2">[2]</a></sup> MRSA, or methicillin-resistant Staphycoccus aureus, and drug-resistant Staphococcus aureus have already <br />
been categorized as “serious threats” by the CDC. <sup><a href="#6">[6]</a></sup>The increasing resistance of these bacteria to the latest generation of antibiotics eventually requires broader spectrum- drugs which have higher costs, higher risks and worse side effects to patients due to a more toxic treatment. The growth of antibiotic resistance in pathogenic bacteria renders one of our most powerful tools against disease useless—making it imperative to develop new courses of treatment.</p><br />
<br />
<p><h4>REFERENCES</h4><br />
<ol><br />
<a id="1"></a> <li> Fleming, Alexander. "Nobel Lecture: Penicillin." Speech, Nobel Prize Award Ceremony from Nobel Foundation, Stockholm, December 11, 1945.</li><br />
<br />
<a id="2"></a><li> World Health Organization. Antimicrobial Resistance: Global Report on Surveillance 2014. S.l.: <br />
<br />
World Health Organization, 2014.</li><br />
<br />
<a id="3"></a><li> Levy, Stuart B, and Bonnie Marshall. "Antibacterial Resistance Worldwide: Causes, Challenges And <br />
<br />
Responses." Nature Medicine 10, no. 12s (2004): S122-S129.</li><br />
<br />
<a id="4"></a><li> Schultz L, Lowe TJ, Srinivasan A, Neilson D, Pugliese G. “Economic Impact of Redundant <br />
<br />
Antimicrobial Therapy in US Hospitals.” Infection Control and Hospital Epidemiliogy 35: 1229-1235.</li><br />
<br />
<a id="5"></a><li>. Rosenblatt-Farrell N. “The Landscape of Antibiotic Resistance.” Environ Health Perspect. 117(6): <br />
<br />
A244-A250.</li><br />
<br />
<a id="6"></a><li> Centers for Disease Control and Prevention. Antibiotic resistance threats in the United States, 2013. <br />
<br />
S.l.: Centers for Disease Control and Prevention, 2013.</li></ol></p><br />
</div><br />
<div class="content-2"><br />
<p> <p><br />
<br />
<h4>OUR MODEL: PSEUDOMONAS AERUGINOSA</h4><br />
<br />
<p>Pseudomonas aeruginosa is an example of an opportunistic and virulent pathogen which is strongly resistant against antibiotics. In addition to its developed antibiotic resistance, P. aeruginosa has intrinsic resistance to antibiotics, due to its low membrane permeability as well as pumps in its membrane which actively pump out antibiotics from the cell.<sup><a href="#8">[1]</a></sup> Multi-drug resistant P. aeruginosa is particularly prevalent in hospitals, where it can colonize in hospital equipment and cause cross-infections in patients with weakened immune systems, such as in patients with cystic fibrosis. <sup><a href="#9">[2]</a></sup> <br />
<br />
<figure id="pa"> <img src="https://static.igem.org/mediawiki/2014/d/dd/Stony_Brook_Project_Paeruginosa.jpg" height="300" width="300"><figcaption> Biofilm formation of P. aeruginosa (<a href="http://www.uoguelph.ca/~confocal/gallery/Image027Snapshot1.jpg"> Source </a>).</figcaption></figure><br />
<br />
Serious P. aeruginosa infections can cause meningitis, pneumonia, and sepsis. <sup><a href="#10">[3]</a></sup> The CDC estimates that 51,000 healthcare-associated infections in the US are caused by P. aeruginosa, 13% of which are multidrug resistant and which the CDC considers a serious threat. <sup><a href="#11">[4]</a></sup> In addition, 30% of healthcare-related P. aeruginosa infections are resistant to fluoroquinolones, a broad-spectrum antibiotic. <sup><a href="#12">[5]</a></sup> The resistance of P. aeruginosa can therefore become deadly for immunocompromised patients, leading to higher mortality rates for affected patients.</p><br />
<br />
<br />
<br />
<h4>THE QUORUM SENSING SYSTEM OF PSEUDOMONAS AERUGINOSA</h4><br />
<br />
<p><br />
The virulence of P. aeruginosa bacteria is in part controlled by their ability to sense each other by producing and recognizing specific compounds called N-acyl homoserine lactones (AHLs), thus allowing them to communicate with each other.</p> <p>This ability to communicate intercellularly, called quorum-sensing, is used by many bacteria to control population growth through gene activation. <figure id="quorumsensing"> <img src="https://static.igem.org/mediawiki/2014/b/b7/Stony_Brook_Project_Quorum_sensing.png"><figcaption> A simple illustration of quorum sensing: Bacteria produce cell-signaling molecules, and at a certain concentration of cells these molecules interact with other bacteria, serving as a communication system </figcaption></figure>As the bacteria grow, they release signaling signals. Once these signals reach a critical concentration, other bacteria in the environment will recognize these signals and respond in turn by making changes in their gene regulation, allowing them to cooperate with each other as their population reaches a certain density. <sup><a href="#13">[6]</a></sup></p><br />
<br />
<br />
<p> P. aeruginosa primarily has two quorum-sensing systems: the las system, which consists of the transcriptional regulator LasR and the synthase protein LasI, and the rhl system, which consists of RhlR and RhlI. LasI and RhlI help in the production of the AHL compounds 3-oxo-C12-HSL and C4-HSL respectively. These compounds bind with their respective transcriptional regulators, LasR or RhlR, which then binds to DNA and regulates the transcription of genes. While both 3-oxo-C12-HSL and C4-HSL freely diffuse out of bacterial cells, 3-oxo-C12-HSL diffuses at a much slower rate than C4-HSL. Both of these systems control the pathogenesis of P.aeruginosa, acting as virulence factors as well as communicative signals and other regulatory factors. <sup><a href="#14">[7]</a></sup></p> <figure class="center"><img src="https://static.igem.org/mediawiki/2014/5/53/Stony_Brook_Project_Rhlr_system.png"><figcaption> As the autoinducer rhli is produced, it forms a complex with C4HSL, which then in turn activates the production of regulatory protein rhlr, which then regulates other genes. </figcaption></figure><br />
<br />
<p><h4>EXPLOITING THE QUORUM-SENSING SYSTEM</h4><br />
<br />
By inserting the genes for the rhl sensing system, non-pathogenic i.e. harmless E. coli can be induced to sense P. aeruginosa signals and act in response to them. If a bacteria-killing compound could be produced upon receiving C4HSL or 3-oxo-C6HSL signals, our project could take advantage of naturally-occurring pathogenic bacterial cell communication in order to kill them off. Since C4-HSL diffuses more quickly, we decided to use the rhl system used by P.aeruginosa as a means of identifying our target bacteria. By exploiting the rhl system in P.aeruginosa, we could <br />
target them with an antibiotic-like compound, thus killing our bacteria.<br />
Our next step was to look into our bacteria-killing compound: melittin, a honeybee venom compound.<br />
</p><br />
<p><h4><br />
REFERENCES</h4><br />
<ol><br />
<a id="8"></a><li> Li XZ, Livermore DM, Nikaido H. 1994. Role of efflux pump(s) in intrinsic resistance of <br />
<br />
Pseudomonas aeruginosa: resistance to tetracycline, chloramphenicol, and norfloxacin. <br />
<br />
Antimicrobial Agents and Chemotherapy. 38:1732-1741</li><br />
<br />
<a id="9"></a><li> Döring G, Parameswaran IG, Murphy TF. 2011. Differential adaptation of microbial <br />
<br />
pathogens to airways of patients with cystic fibrosis and chronic obstructive pulmonary <br />
<br />
disease. FEMS Microbiol. Rev. 35:124–146</li><br />
<br />
<a id="10"></a><li> Bodey GP, Bolivar R, Fainstein V, Jadeja L. 1983. Infections caused by Pseudomonas <br />
<br />
aeruginosa. Rev Infect Dis. 5:279-313.</li><br />
<br />
<a id="11"></a><li> "Pseudomonas aeruginosa in Healthcare Settings." Centers for Disease Control and <br />
<br />
Prevention. Centers for Disease Control and Prevention, 7 May 2014. Web. 3 June 2014.</li><br />
<br />
<a id="12"></a><li> Septimus EJ, Kuper KM. 2009. Clinical challenges in addressing resistance to <br />
<br />
antimicrobial drugs in the twenty-first century. Clin Pharmacol Ther. 86(3): 336-339.</li><br />
<br />
<br />
<a id="13"></a><li> Difri, CD. 2008. Difri, CD. 2008. Quorum Sensing: Bacteria Talk Sense. Clin Infect Dis. 47 (8): 1070-1076</li><br />
<br />
<br />
<a id="14"></a><li> Smith RS, Iglewski BH. 2003. Pseudomonas aeruginosa quorum sensing as a potential <br />
<br />
antimicrobial target. J Clin Invest. 112(10): 1460-1465</li></ol></p> </p><br />
</div><br />
<div class="content-3"><br />
<h4>HONEYBEE VENOM: PREPROMELITTIN AND MELITTIN</h4><br />
<p>Honeybees, or Apis mellifera, are native to Europe, Africa, and western Asia. The introduction of Apis mellifera to other parts of the world began in the 17th century and since then, honeybees can be found around the world. Currently, there are 26 recognized subspecies of Apis mellifera based on morphology and molecular differences, as well as the differences in their habitat.While male bee drones do not have stingers, female bees have a stinger, which acts as a form of defense against predation. It is connected to a sac that releases a venom when the bee stings another organism. <figure id="bee"> <img src="https://static.igem.org/mediawiki/2014/e/e4/Stony_Brook_Project_Apismellifera.jpg" height="300" width="400"> <figcaption> Apis mellifera( <a href="http://en.wikipedia.org/wiki/Italian_bee#mediaviewer/File:Honeybee-27527-1.jpg"> Source </a>) </figcaption></figure></p><br />
<p>Bee venom has been researched for its ability to treat inflammation, infections, and auto-immune diseases. The active portion of the venom causes inflammation and acts as an anticoagulant. The primary active component of bee venom is melittin, which constitutes about 52% of the apitoxin, or bee venom, liquid excreted from the bee. <sup><a href="#15">[1]</a></sup> An antimicrobial peptide, melittin is a part of the host defense system and immune response of honeybees. Previous research has shown it to be effective with some modifications in both treating cancerous cells and HIV.<sup><a href="#16">[2]</a></sup><sup><a href="#17">[3]</a></sup></p><br />
<br />
<p>Melittin is not produced in bees in its active form. Rather, it is produced in an inactivated form called prepromelittin, which contains the right sequences to send it to the endoplasmic reticulum in bee cells as well as deactivating it. <br />
Sequence prepromelittin<br />
Naturally, bees produce prepromelittin in their venom gland, which gets degraded by surrounding enzymes. <sup> <a href="#18">[4]</a></sup> As the prepromelittin is processed in the honeybee cell, the “pre” part of the peptide, which acts as a signal peptide, is cleaved off by signal peptidase enzymes. <sup> <a href="#19">[5]</a></sup> The “pro” sequence, which acts as a protective sequence which inactivates melittin, is slowly cleaved off stepwise by surrounding dipeptidase enzymes as well, allowing the melittin to become liberated.<sup><a href="#20">[6]</a></sup></p><br />
<br />
<figure class="center"><img src="https://static.igem.org/mediawiki/2014/1/1a/Stony_Brook_Project_Prepromelittin_sequence.png"><figcaption> Sequence of prepromelittin, where green indicates "pre" sequence, red indicated "pro" sequence, and blue indicates the melittin sequence.</figcaption></figure><br />
<br />
<h4>MELITTIN IN ACTION </h4><br />
<p>Melittin is a small ampipathic peptide made up of 26 amino acids, with a hydrophobic N-terminus and a hydrophilic C-terminus.As melittin concentration in the membrane accumulates, transient pores are formed which allow the permeability of ions. <figure id="melittin"><img src="https://static.igem.org/mediawiki/2014/0/0c/Stony_Brook_Project_Melittin.jpg"><figcaption>Melittin's alpha-helical structure.</figcaption></figure>As the concentration of melittin increases further, pores are stabilized and thus become large enough to allow the movement of large molecules outside the cell, and at even higher concentrations, the membrane will disintegrate entirely. Effectively, the melittin compound will cause cells to lyse either due to osmotic pressure around the cell, or through a detergent-effect at high enough concentrations. <br />
Its activity as a cytolytic compound is due to its surface activity. At low concentrations, it adopts an inactive parallel orientation to the cell membrane, and at higher concentrations it adopts a perpendicular orientation to the membrane, which allows it to “wedge” itself in the headgroup space of phospholipids in a phospholipid bilayer, without extending all the way to the center of the bilayer. As a result, the bilayer distorts and curves to fill in the space left by the “wedge”, forming transient pores through the resulting area imbalance between the inner and outer part of the membrane. Through the transient pores, melittin molecules redistribute, eventually forming larger stable pores made up of 4-7 melittin monomers each.<sup><a href="#21">[7]</a></sup> </p><br />
<br />
<h4>WHY USE MELITTIN?</h4><br />
<p><figure id="pores"> <img src="https://static.igem.org/mediawiki/2014/7/77/Stony_Brook_Project_Melittin_pore_formation_mechanism.jpg"><figcaption> Mechanism of pore formation (<a href="http://neutron.kaist.ac.kr/research/poreformation.jpg"> Source </a>) </figcaption></figure>Since the majority of antibiotics target parts of bacteria which have mutated to develop resistance, the most effective alternative to antibiotics would work against conserved parts of the bacteria, which cannot mutate easily without compromising its ability to thrive. An example of such an area would be the outermost part of the bacteria, the cell membrane. Melittin can work against pathogenic bacteria by targeting their cell membrane. Thus, melittin can be utilized against pathogenic bacteria as an alternative to antibiotics, as well as a solution to the increasing resistance of bacteria against antibiotics.</p><br />
<br />
<h4> REFERENCES</h4><br />
<br />
<ol><a id="15"></a><li>Meier J, White J. (1995). Clinical toxicology of animal venoms and poisons. CRC Press, Inc. ISBN 0-8493-4489-1.</li><br />
<a id="16"></a><li>Soman NR, Baldwin SL, Hu G, Marsh JN, Lanza GM, Heuser JE, Arbeit JM, Wickline SA, Schlesinger PH. 2009. Molecularly targeted nanocarriers deliver the cytolytic peptide specifically to tumor cells in mice, reduing cancer growth. J Clin Invest. 199(9): 2830-2842</li><br />
<a id="17"></a><li>Hood JL, Jallouk AP, Campbell N, Ratner L, Wickline SA. 2013. Cytolytic nanoparticles attenuate HIC-1 infectivity. Antivi Ther. 18(1):95-103</li><br />
<a id="18"></a><li>Müller G, Zimmermann R. 1987. Import of honeybee prepromelittin into the endoplasmic reticulum: structural basis for independence of SRP and docking protein. EMBO J. 6:2099-2107</li><br />
<a id="19"></a><li>Mollay C, Vilas U, Kreil G. 1982. Cleavage of honeybee prepromelittin by an endoprotease from rat liver microsomes: identification of intact signal peptide. Proc Natl Acad Sci USA. 79(7): 2260-2263</li><br />
<a id="20"></a><li>Kreil G, Haiml L, Suchanek G. 1980. Stepwise cleavage of the pro part of promelittin by dipeptidylpeptidase IV. Evidence for a new type of precursor-product conversion. Eur J Biochem. 111(1):49-58 </li><br />
<a id="21"></a><li>Lee MT, Sun TL, Hung WC, Huang HW. 2013. Process of inducing pores in membranes by melittin. Proc Natl Acad Sci USA. 110(35): 14243-8</li></ol><br />
<br />
</div><br />
<div class="content-4"><br />
<br />
<br />
<br />
<br />
<h4> GST-MELITTIN </h4> <figure class="center"><img src="https://static.igem.org/mediawiki/2014/7/7f/Stony_Brook_Project_GST-Melittin.png"> </figure><p>In order to prevent melittin from either attacking our own cells or from congregating into inactive tetramers, melittin itself cannot be produced within our E.coli cells. However, if we were to produce the naturally-occurring form of prepromelittin, we would also need to recreate the enzymes within the bee. Reproducing these enzymes in an E. coli cell can quickly become complicated, therefore our project uses the GST tag to inactivate melittin, creating a fusion protein. This tag also allows for easy identification by through gel electrophoresis, allowing us to also see if the melittin was produced. A TEV protease site sits in between the GST tag and melittin, allowing for easy cleavage of the GST tag and activation of melittin upon the addition of the TEV protease.</p><br />
<h4>RHLR BIOSENSOR</h4><br />
<figure class="center"><img src="https://static.igem.org/mediawiki/2014/6/6b/Stony_Brook_Project_RhlR.png"></figure><p>We designed a biosensor which could detect the quorum sensing signals of P. aeruginosa. This biosensor would constitutively produce the RhlR regulatory protein in order to bind with C4HSL molecules, which then would activate the fluorescence marker mCherry. By responding to C4HSL molecules in the environment, our biosensor could then activate the production of our antibacterial compound, melittin. This form of mechanism allows for a more fine-tuned production of melittin, and acts as a preliminary check for biosafety and the efficiency of our circuit.</p><br />
<h4>OUR COMPLETE CIRCUIT(RHLR+MELITTIN)</h4><br />
<br/><br />
<br/><br />
<figure class="center"><img src="https://static.igem.org/mediawiki/2014/4/47/Stony_Brook_Project_Entire_circuit.png"></figure><br />
<p> In our complete circuit, our <i>E. coli</i> would constitutively produce RhlR proteins, which then could bind with any C4HSL signal molecules in the environment released by P. aeruginosa. Then, this would activate the <i>rhl</i> promoter, starting the production of GST-melittin. We then would exogenously add in TEV protease, in order to cleave off the GST protein and allow the melittin to work against our target bacteria. Both the RhlR and GST-melittin have fluorescence markers, to allow us to easily monitor the progress of the mechanism. In addition, the use of non-specific melittin also acts as a "kill switch", as our <i>E.coli</i> would eventually succumb to the pores created by melittin. </p><br />
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Hliu1423
http://2014.igem.org/Team:Stony_Brook/Project
Team:Stony Brook/Project
2014-10-18T01:26:06Z
<p>Hliu1423: </p>
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<div id="yellow_header"> <a href="http://www.stonybrook.edu/">Stony Brook University</a></div><br />
<br />
<div id="blue_header"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/1/1e/Stony_Brook_Apis-biotics_header.png"/ id="apis-biotics"/></a><br />
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<p> Attributions </p><br />
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<br />
<div id="project"><p>Project</p> <br />
</div><br />
<br />
<div id="abstract"> <h3>Abstract</h3><br />
<p>Pathogenic bacteria are becoming increasingly antibiotic-resistant due to misuse, overuse and abuse. In addition, infections caused by some harmful strains of bacteria, particularly gram-negative bacteria, cannot easily be treated with antibiotics or other common forms of treatment. Our intention is to engineer cells which can both produce an antimicrobial peptide and recognize the communication signals of such bacteria, using Pseudomonas aeruginosa as a model. Two plasmids, one which controls the production of our antimicrobial peptide melittin and the other which acts as the cell-signal receiver, will be inserted in nonpathogenic E. coli cells. This allows our E. coli to recognize the cells of Pseudomonas aeruginosa, and release a bacteria-killing compound in response. </p><br />
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<div id="slideshow"><img src="https://static.igem.org/mediawiki/2014/a/ad/Stony_Brook_Project_Slideshow1.png" height="100%"></div><br />
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<div id="contents"><br />
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<label for="tab-1" class="tab-label-1">Antibiotic resistance</label><br />
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<label for="tab-2" class="tab-label-2">P. aeruginosa</label><br />
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<label for="tab-3" class="tab-label-3">Melittin</label><br />
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<label for="tab-4" class="tab-label-4">Our Solution: Apis-Biotics</label><br />
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<p><br />
<h4>WHEN ANTIBIOTICS FAIL</h4><br />
<br />
When Alexander Fleming was accepting his Nobel Prize in medicine for his discovery of penicillin, the world’s first antibiotic, he warned of a future where antibiotics are no longer effective. <sup><a href="#1">[1]</a></sup> The growth of antibacterial resistance—when bacteria no longer are affected by antibiotics—is quickly becoming a major public health concern. In 2014, the World Health Organization stated “A post-antibiotic era—in which common infections and minor injuries can kill—far from being an apocalyptic fantasy, is instead a very real possibility for the 21<sup>st</sup> century.” <sup><a href="#2">[2]</a></sup> </p><br />
<br />
<h4>THE MECHANISM OF ANTIBIOTIC RESISTANCE</h4><br />
<br />
<p>Antibiotic resistance is a natural phenomenon that occurs when mutations in bacterial replications render antibiotics ineffective. Many antibiotics kill bacteria by either preventing <br />
them from creating bacterial cell walls, affecting their DNA, RNA, or proteins, or changing their metabolism. However, through mutations, bacteria may develop the ability to break down <figure id="betalactamase"><br />
<img src="https://static.igem.org/mediawiki/2014/c/c9/Stony_Brook_Project_Beta_lactam_penicillin_action.png"><br />
<figcaption>Bacteria are constantly rebuilding their cell walls through peptidoglycan synthesis. Penicillin works by deactivating transpeptidase within the bacteria, preventing the cross-linking of peptidoglycan and weakening the cell wall, eventually causing lysis due to osmotic pressure. Bacteria can develop resistance to penicillin by producing β-lactamase, which breaks the beta-lactam ring in penicillin, deactivating it.</figcaption></figure>these antibiotics, prevent the antibiotic from entering the cell, or use efflux pumps in the cell membrane to transport the antibiotic back outside the cell. When antibiotics are used to kill off harmful bacteria, the resistant strain of bacteria remains and reproduces. With other strains <br />
gone, this resistant strain has less competition, allowing it to thrive.<br />
<br />
</p><br />
<br />
<p>However, what occurs more often than mutation-derived resistance is the spread of antibiotic resistance through horizontal gene transfer, which occurs when genetic material from bacteria, such as plasmids, transposons, or DNA are passed from cell to cell. Through this mechanism, bacteria can even develop multi-drug resistance. Even when a singular antibiotic is used for more than ten days, multidrug resistance develops to structurally unrelated drugs as the resistant bacteria recruit resistance genes from other bacteria in the environment, encouraging the development of “super-bugs.” In addition, some bacteria are intrinsically resistant to bacteria, such as gram-negative bacteria, which due to its thicker outer cell membrane are harder to treat with certain classes of antibiotics. In some cases, some strains of gram-negative bacteria are resistant to all antibiotics, even the <br />
newest families of antibiotics and antibiotics used as a last resort. <sup><a href="#3">[3]</a></sup><br />
<figure id="antibioticresistance"><img src="https://static.igem.org/mediawiki/2014/c/ce/Stony_Brook_Project_Antibiotic_resistance_selection.png"><figcaption>After an antibiotic is used, bacteria with resistance genes are selected while those without resistance die off, allowing resistant bacteria to better compete for resources and thrive</figcaption></figure><figure id="horizontal"><img src="https://static.igem.org/mediawiki/2014/4/43/Stony_Brook_Project_Horizontal_gene_transfer.png"><figcaption> Horizontal gene transfer: DNA for antibiotic resistance is transfered through conjugation.</figcaption></figure><br />
</p><br />
<br />
<br />
<p>The misuse of antibiotics in the healthcare environment contributes one avenue for the spread of antibiotic resistance. In one study done for the CDC, researchers found that almost 80% of hospitals misuse antibiotics, either giving inappropriate or redundant antibiotic combinations and intravenous antibiotics. <sup><a href="#4">[4]</a></sup> The use of antibiotics in food and agriculture also presents a significant problem. The Union of Concerned Scientists estimated in one report that 70% of all antibiotics used in the U.S. are used in the food and water of already-healthy livestock, in order to control disease. Many of these antibiotics don’t break down in the waste of livestock, leading to the growth of resistant bacteria which spreads as this waste is used as fertilizer, contaminating the soil and sources of water. This contamination can also be found coming from antibiotic manufacturing plants, as well as households which improperly dispose of unused or expired <br />
antibiotic pills, instead or returning them to a pharmacy. <sup><a href="#4">[5]</a></sup></p><br />
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<h4>TO WHAT EXTENT?</h4><br />
<br />
<p>In its 2014 report on antibiotic resistance, the World Health Organization reported the current status of antibiotic resistance in seven major bacteria which cause disease in humans, including Escherichia coli, Klebsiella pneumonia, Staphylococcus aureus, and Streptococcus pneumoniae. These bacteria are common causes of infections within a hospital or community. If they become highly resistant to antibiotics, there would be significant repercussions for public health <br />
worldwide. </p><br />
<br />
<figure class="center"><img src='https://static.igem.org/mediawiki/2014/b/bc/Stony_Brook_Project_Antibiotic_resistance_chart.png'><figcaption> Data taken from WHO 2014 report on antimicrobial resistance </figcaption></figure><br />
<br />
<h4>WHY DOES IT MATTER?</h4><br />
<br />
<p>Antibiotic resistance is a global concern—it jeopardizes our ability to treat common infections, causing standard treatments to be replaced by more expensive drug therapies, making infectious diseases longer to treat and easier to spread, putting major surgeries, chemotherapy and organ transplants at a greater risk of being unsuccessful, and increasing death rate. <sup><a href="#2">[2]</a></sup> MRSA, or methicillin-resistant Staphycoccus aureus, and drug-resistant Staphococcus aureus have already <br />
been categorized as “serious threats” by the CDC. <sup><a href="#6">[6]</a></sup>The increasing resistance of these bacteria to the latest generation of antibiotics eventually requires broader spectrum- drugs which have higher costs, higher risks and worse side effects to patients due to a more toxic treatment. The growth of antibiotic resistance in pathogenic bacteria renders one of our most powerful tools against disease useless—making it imperative to develop new courses of treatment.</p><br />
<br />
<p><h4>REFERENCES</h4><br />
<ol><br />
<a id="1"></a> <li> Fleming, Alexander. "Nobel Lecture: Penicillin." Speech, Nobel Prize Award Ceremony from Nobel Foundation, Stockholm, December 11, 1945.</li><br />
<br />
<a id="2"></a><li> World Health Organization. Antimicrobial Resistance: Global Report on Surveillance 2014. S.l.: <br />
<br />
World Health Organization, 2014.</li><br />
<br />
<a id="3"></a><li> Levy, Stuart B, and Bonnie Marshall. "Antibacterial Resistance Worldwide: Causes, Challenges And <br />
<br />
Responses." Nature Medicine 10, no. 12s (2004): S122-S129.</li><br />
<br />
<a id="4"></a><li> Schultz L, Lowe TJ, Srinivasan A, Neilson D, Pugliese G. “Economic Impact of Redundant <br />
<br />
Antimicrobial Therapy in US Hospitals.” Infection Control and Hospital Epidemiliogy 35: 1229-1235.</li><br />
<br />
<a id="5"></a><li>. Rosenblatt-Farrell N. “The Landscape of Antibiotic Resistance.” Environ Health Perspect. 117(6): <br />
<br />
A244-A250.</li><br />
<br />
<a id="6"></a><li> Centers for Disease Control and Prevention. Antibiotic resistance threats in the United States, 2013. <br />
<br />
S.l.: Centers for Disease Control and Prevention, 2013.</li></ol></p><br />
</div><br />
<div class="content-2"><br />
<p> <p><br />
<br />
<h4>OUR MODEL: PSEUDOMONAS AERUGINOSA</h4><br />
<br />
<p>Pseudomonas aeruginosa is an example of an opportunistic and virulent pathogen which is strongly resistant against antibiotics. In addition to its developed antibiotic resistance, P. aeruginosa has intrinsic resistance to antibiotics, due to its low membrane permeability as well as pumps in its membrane which actively pump out antibiotics from the cell.<sup><a href="#8">[1]</a></sup> Multi-drug resistant P. aeruginosa is particularly prevalent in hospitals, where it can colonize in hospital equipment and cause cross-infections in patients with weakened immune systems, such as in patients with cystic fibrosis. <sup><a href="#9">[2]</a></sup> <br />
<br />
<figure id="pa"> <img src="https://static.igem.org/mediawiki/2014/d/dd/Stony_Brook_Project_Paeruginosa.jpg" height="300" width="300"><figcaption> Biofilm formation of P. aeruginosa (<a href="http://www.uoguelph.ca/~confocal/gallery/Image027Snapshot1.jpg"> Source </a>).</figcaption></figure><br />
<br />
Serious P. aeruginosa infections can cause meningitis, pneumonia, and sepsis. <sup><a href="#10">[3]</a></sup> The CDC estimates that 51,000 healthcare-associated infections in the US are caused by P. aeruginosa, 13% of which are multidrug resistant and which the CDC considers a serious threat. <sup><a href="#11">[4]</a></sup> In addition, 30% of healthcare-related P. aeruginosa infections are resistant to fluoroquinolones, a broad-spectrum antibiotic. <sup><a href="#12">[5]</a></sup> The resistance of P. aeruginosa can therefore become deadly for immunocompromised patients, leading to higher mortality rates for affected patients.</p><br />
<br />
<br />
<br />
<h4>THE QUORUM SENSING SYSTEM OF PSEUDOMONAS AERUGINOSA</h4><br />
<br />
<p><br />
The virulence of P. aeruginosa bacteria is in part controlled by their ability to sense each other by producing and recognizing specific compounds called N-acyl homoserine lactones (AHLs), thus allowing them to communicate with each other.</p> <p>This ability to communicate intercellularly, called quorum-sensing, is used by many bacteria to control population growth through gene activation. <figure id="quorumsensing"> <img src="https://static.igem.org/mediawiki/2014/b/b7/Stony_Brook_Project_Quorum_sensing.png"><figcaption> A simple illustration of quorum sensing: Bacteria produce cell-signaling molecules, and at a certain concentration of cells these molecules interact with other bacteria, serving as a communication system </figcaption></figure>As the bacteria grow, they release signaling signals. Once these signals reach a critical concentration, other bacteria in the environment will recognize these signals and respond in turn by making changes in their gene regulation, allowing them to cooperate with each other as their population reaches a certain density. <sup><a href="#13">[6]</a></sup></p><br />
<br />
<br />
<p> P. aeruginosa primarily has two quorum-sensing systems: the las system, which consists of the transcriptional regulator LasR and the synthase protein LasI, and the rhl system, which consists of RhlR and RhlI. LasI and RhlI help in the production of the AHL compounds 3-oxo-C12-HSL and C4-HSL respectively. These compounds bind with their respective transcriptional regulators, LasR or RhlR, which then binds to DNA and regulates the transcription of genes. While both 3-oxo-C12-HSL and C4-HSL freely diffuse out of bacterial cells, 3-oxo-C12-HSL diffuses at a much slower rate than C4-HSL. Both of these systems control the pathogenesis of P.aeruginosa, acting as virulence factors as well as communicative signals and other regulatory factors. <sup><a href="#14">[7]</a></sup></p> <figure class="center"><img src="https://static.igem.org/mediawiki/2014/5/53/Stony_Brook_Project_Rhlr_system.png"><figcaption> As the autoinducer rhli is produced, it forms a complex with C4HSL, which then in turn activates the production of regulatory protein rhlr, which then regulates other genes. </figcaption></figure><br />
<br />
<p><h4>EXPLOITING THE QUORUM-SENSING SYSTEM</h4><br />
<br />
By inserting the genes for the rhl sensing system, non-pathogenic i.e. harmless E. coli can be induced to sense P. aeruginosa signals and act in response to them. If a bacteria-killing compound could be produced upon receiving C4HSL or 3-oxo-C6HSL signals, our project could take advantage of naturally-occurring pathogenic bacterial cell communication in order to kill them off. Since C4-HSL diffuses more quickly, we decided to use the rhl system used by P.aeruginosa as a means of identifying our target bacteria. By exploiting the rhl system in P.aeruginosa, we could <br />
target them with an antibiotic-like compound, thus killing our bacteria.<br />
Our next step was to look into our bacteria-killing compound: melittin, a honeybee venom compound.<br />
</p><br />
<p><h4><br />
REFERENCES</h4><br />
<ol><br />
<a id="8"></a><li> Li XZ, Livermore DM, Nikaido H. 1994. Role of efflux pump(s) in intrinsic resistance of <br />
<br />
Pseudomonas aeruginosa: resistance to tetracycline, chloramphenicol, and norfloxacin. <br />
<br />
Antimicrobial Agents and Chemotherapy. 38:1732-1741</li><br />
<br />
<a id="9"></a><li> Döring G, Parameswaran IG, Murphy TF. 2011. Differential adaptation of microbial <br />
<br />
pathogens to airways of patients with cystic fibrosis and chronic obstructive pulmonary <br />
<br />
disease. FEMS Microbiol. Rev. 35:124–146</li><br />
<br />
<a id="10"></a><li> Bodey GP, Bolivar R, Fainstein V, Jadeja L. 1983. Infections caused by Pseudomonas <br />
<br />
aeruginosa. Rev Infect Dis. 5:279-313.</li><br />
<br />
<a id="11"></a><li> "Pseudomonas aeruginosa in Healthcare Settings." Centers for Disease Control and <br />
<br />
Prevention. Centers for Disease Control and Prevention, 7 May 2014. Web. 3 June 2014.</li><br />
<br />
<a id="12"></a><li> Septimus EJ, Kuper KM. 2009. Clinical challenges in addressing resistance to <br />
<br />
antimicrobial drugs in the twenty-first century. Clin Pharmacol Ther. 86(3): 336-339.</li><br />
<br />
<br />
<a id="13"></a><li> Difri, CD. 2008. Difri, CD. 2008. Quorum Sensing: Bacteria Talk Sense. Clin Infect Dis. 47 (8): 1070-1076</li><br />
<br />
<br />
<a id="14"></a><li> Smith RS, Iglewski BH. 2003. Pseudomonas aeruginosa quorum sensing as a potential <br />
<br />
antimicrobial target. J Clin Invest. 112(10): 1460-1465</li></ol></p> </p><br />
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<div class="content-3"><br />
<h4>HONEYBEE VENOM: PREPROMELITTIN AND MELITTIN</h4><br />
<p>Honeybees, or Apis mellifera, are native to Europe, Africa, and western Asia. The introduction of Apis mellifera to other parts of the world began in the 17th century and since then, honeybees can be found around the world. Currently, there are 26 recognized subspecies of Apis mellifera based on morphology and molecular differences, as well as the differences in their habitat.While male bee drones do not have stingers, female bees have a stinger, which acts as a form of defense against predation. It is connected to a sac that releases a venom when the bee stings another organism. <figure id="bee"> <img src="https://static.igem.org/mediawiki/2014/e/e4/Stony_Brook_Project_Apismellifera.jpg" height="300" width="400"> <figcaption> Apis mellifera( <a href="http://en.wikipedia.org/wiki/Italian_bee#mediaviewer/File:Honeybee-27527-1.jpg"> Source </a>) </figcaption></figure></p><br />
<p>Bee venom has been researched for its ability to treat inflammation, infections, and auto-immune diseases. The active portion of the venom causes inflammation and acts as an anticoagulant. The primary active component of bee venom is melittin, which constitutes about 52% of the apitoxin, or bee venom, liquid excreted from the bee. <sup><a href="#15">[1]</a></sup> An antimicrobial peptide, melittin is a part of the host defense system and immune response of honeybees. Previous research has shown it to be effective with some modifications in both treating cancerous cells and HIV.<sup><a href="#16">[2]</a></sup><sup><a href="#17">[3]</a></sup></p><br />
<br />
<p>Melittin is not produced in bees in its active form. Rather, it is produced in an inactivated form called prepromelittin, which contains the right sequences to send it to the endoplasmic reticulum in bee cells as well as deactivating it. <br />
Sequence prepromelittin<br />
Naturally, bees produce prepromelittin in their venom gland, which gets degraded by surrounding enzymes. <sup> <a href="#18">[4]</a></sup> As the prepromelittin is processed in the honeybee cell, the “pre” part of the peptide, which acts as a signal peptide, is cleaved off by signal peptidase enzymes. <sup> <a href="#19">[5]</a></sup> The “pro” sequence, which acts as a protective sequence which inactivates melittin, is slowly cleaved off stepwise by surrounding dipeptidase enzymes as well, allowing the melittin to become liberated.<sup><a href="#20">[6]</a></sup></p><br />
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<figure class="center"><img src="https://static.igem.org/mediawiki/2014/1/1a/Stony_Brook_Project_Prepromelittin_sequence.png"><figcaption> Sequence of prepromelittin, where green indicates "pre" sequence, red indicated "pro" sequence, and blue indicates the melittin sequence.</figcaption></figure><br />
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<h4>MELITTIN IN ACTION </h4><br />
<p>Melittin is a small ampipathic peptide made up of 26 amino acids, with a hydrophobic N-terminus and a hydrophilic C-terminus.As melittin concentration in the membrane accumulates, transient pores are formed which allow the permeability of ions. <figure id="melittin"><img src="https://static.igem.org/mediawiki/2014/0/0c/Stony_Brook_Project_Melittin.jpg"><figcaption>Melittin's alpha-helical structure.</figcaption></figure>As the concentration of melittin increases further, pores are stabilized and thus become large enough to allow the movement of large molecules outside the cell, and at even higher concentrations, the membrane will disintegrate entirely. Effectively, the melittin compound will cause cells to lyse either due to osmotic pressure around the cell, or through a detergent-effect at high enough concentrations. <br />
Its activity as a cytolytic compound is due to its surface activity. At low concentrations, it adopts an inactive parallel orientation to the cell membrane, and at higher concentrations it adopts a perpendicular orientation to the membrane, which allows it to “wedge” itself in the headgroup space of phospholipids in a phospholipid bilayer, without extending all the way to the center of the bilayer. As a result, the bilayer distorts and curves to fill in the space left by the “wedge”, forming transient pores through the resulting area imbalance between the inner and outer part of the membrane. Through the transient pores, melittin molecules redistribute, eventually forming larger stable pores made up of 4-7 melittin monomers each.<sup><a href="#21">[7]</a></sup> </p><br />
<br />
<h4>WHY USE MELITTIN?</h4><br />
<p><figure id="pores"> <img src="https://static.igem.org/mediawiki/2014/7/77/Stony_Brook_Project_Melittin_pore_formation_mechanism.jpg"><figcaption> Mechanism of pore formation (<a href="http://neutron.kaist.ac.kr/research/poreformation.jpg"> Source </a>) </figcaption></figure>Since the majority of antibiotics target parts of bacteria which have mutated to develop resistance, the most effective alternative to antibiotics would work against conserved parts of the bacteria, which cannot mutate easily without compromising its ability to thrive. An example of such an area would be the outermost part of the bacteria, the cell membrane. Melittin can work against pathogenic bacteria by targeting their cell membrane. Thus, melittin can be utilized against pathogenic bacteria as an alternative to antibiotics, as well as a solution to the increasing resistance of bacteria against antibiotics.</p><br />
<br />
<h4> REFERENCES</h4><br />
<br />
<ol><a id="15"></a><li>Meier J, White J. (1995). Clinical toxicology of animal venoms and poisons. CRC Press, Inc. ISBN 0-8493-4489-1.</li><br />
<a id="16"></a><li>Soman NR, Baldwin SL, Hu G, Marsh JN, Lanza GM, Heuser JE, Arbeit JM, Wickline SA, Schlesinger PH. 2009. Molecularly targeted nanocarriers deliver the cytolytic peptide specifically to tumor cells in mice, reduing cancer growth. J Clin Invest. 199(9): 2830-2842</li><br />
<a id="17"></a><li>Hood JL, Jallouk AP, Campbell N, Ratner L, Wickline SA. 2013. Cytolytic nanoparticles attenuate HIC-1 infectivity. Antivi Ther. 18(1):95-103</li><br />
<a id="18"></a><li>Müller G, Zimmermann R. 1987. Import of honeybee prepromelittin into the endoplasmic reticulum: structural basis for independence of SRP and docking protein. EMBO J. 6:2099-2107</li><br />
<a id="19"></a><li>Mollay C, Vilas U, Kreil G. 1982. Cleavage of honeybee prepromelittin by an endoprotease from rat liver microsomes: identification of intact signal peptide. Proc Natl Acad Sci USA. 79(7): 2260-2263</li><br />
<a id="20"></a><li>Kreil G, Haiml L, Suchanek G. 1980. Stepwise cleavage of the pro part of promelittin by dipeptidylpeptidase IV. Evidence for a new type of precursor-product conversion. Eur J Biochem. 111(1):49-58 </li><br />
<a id="21"></a><li>Lee MT, Sun TL, Hung WC, Huang HW. 2013. Process of inducing pores in membranes by melittin. Proc Natl Acad Sci USA. 110(35): 14243-8</li></ol><br />
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<h4> GST-MELITTIN </h4> <figure class="center"><img src="https://static.igem.org/mediawiki/2014/7/7f/Stony_Brook_Project_GST-Melittin.png"> </figure><p>In order to prevent melittin from either attacking our own cells or from congregating into inactive tetramers, melittin itself cannot be produced within our E.coli cells. However, if we were to produce the naturally-occurring form of prepromelittin, we would also need to recreate the enzymes within the bee. Reproducing these enzymes in an E. coli cell can quickly become complicated, therefore our project uses the GST tag to inactivate melittin, creating a fusion protein. This tag also allows for easy identification by through gel electrophoresis, allowing us to also see if the melittin was produced. A TEV protease site sits in between the GST tag and melittin, allowing for easy cleavage of the GST tag and activation of melittin upon the addition of the TEV protease.</p><br />
<h4>RHLR BIOSENSOR</h4><br />
<figure class="center"><img src="https://static.igem.org/mediawiki/2014/6/6b/Stony_Brook_Project_RhlR.png"></figure><p>We designed a biosensor which could detect the quorum sensing signals of P. aeruginosa. This biosensor would constitutively produce the RhlR regulatory protein in order to bind with C4HSL molecules, which then would activate the fluorescence marker mCherry. By responding to C4HSL molecules in the environment, our biosensor could then activate the production of our antibacterial compound, melittin. This form of mechanism allows for a more fine-tuned production of melittin, and acts as a preliminary check for biosafety and the efficiency of our circuit.</p><br />
<h4>OUR COMPLETE CIRCUIT(RHLR+MELITTIN)</h4><br />
<br/><br />
<br/><br />
<figure class="center"><img src="https://static.igem.org/mediawiki/2014/4/47/Stony_Brook_Project_Entire_circuit.png"></figure><br />
<p> In our complete circuit, our <i>E. coli</i> would constitutively produce RhlR proteins, which then could bind with any C4HSL signal molecules in the environment released by P. aeruginosa. Then, this would activate the <i>rhl</i> promoter, starting the production of GST-melittin. We then would exogenously add in TEV protease, in order to cleave off the GST protein and allow the melittin to work against our target bacteria. Both the RhlR and GST-melittin have fluorescence markers, to allow us to easily monitor the progress of the mechanism. In addition, the use of non-specific melittin also acts as a "kill switch", as our <i>E.coli</i> would eventually succumb to the pores created by melittin. </p><br />
<br />
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Hliu1423
http://2014.igem.org/Team:Stony_Brook
Team:Stony Brook
2014-10-18T00:45:02Z
<p>Hliu1423: </p>
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<h1> Project Description </h1><br />
<p> The threat of antibiotic resistance has been a concern since the discovery of penicillin. As antibiotics continue to be used to treat disease, pathogenic bacteria continue to develop resistance, eventually becoming irresponsive to multiple classes of antibiotics. This is a major problem for areas that must maintain sterile environments, such as hospitals or other health-care settings. Hospital-acquired infections (HAI) are already an existing issue that poses risks to people receiving or recovering from treatment. Gram-negative bacteria in particular are known to cause more HAIs as they are more resistant to antibiotics than gram-positive bacteria. Irresponsible use of antibiotics has only exacerbated this issue. Recent reports from the World Health Organization confirm that antibiotic resistance is no longer something to worry about in the future, but a current, global health threat. </p><br />
<br />
<p> Alternatives to modern antibiotics, such as antimicrobial peptides, are now being researched. Antimicrobial peptides, or AMPs, are peptides produced innately by the immune systems of certain organisms to fight off infection. Because they kill microorganisms through different mechanisms, they have the potential to be effective against antibiotic resistant bacteria. One such AMP is melittin, produced by honey bees and used in their venom. Melittin kills cells by forming pores in the cell membrane, eventually destabilizing the cell and causing cell lysis. Because melittin targets a conserved area of cells, bacteria cannot develop resistance to it as easily as different types of antibiotics. </p><br />
<br />
<p> Our project this summer is to transform E. coli with the melittin gene in honey bees, so that the E. coli may express the melittin peptide, thereby developing a means to mass-produce melittin as an alternative to today's antibiotics. We hope to optimize the production of melittin in E. coli, as well as to test the bioactivity of our produced melittin in comparison to its naturally-occurring counterpart. </p><br />
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Hliu1423
http://2014.igem.org/Team:Stony_Brook
Team:Stony Brook
2014-10-18T00:44:03Z
<p>Hliu1423: </p>
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<!-- <div id="projectDescription"><br />
<h1> Project Description </h1><br />
<p> The threat of antibiotic resistance has been a concern since the discovery of penicillin. As antibiotics continue to be used to treat disease, pathogenic bacteria continue to develop resistance, eventually becoming irresponsive to multiple classes of antibiotics. This is a major problem for areas that must maintain sterile environments, such as hospitals or other health-care settings. Hospital-acquired infections (HAI) are already an existing issue that poses risks to people receiving or recovering from treatment. Gram-negative bacteria in particular are known to cause more HAIs as they are more resistant to antibiotics than gram-positive bacteria. Irresponsible use of antibiotics has only exacerbated this issue. Recent reports from the World Health Organization confirm that antibiotic resistance is no longer something to worry about in the future, but a current, global health threat. </p><br />
<br />
<p> Alternatives to modern antibiotics, such as antimicrobial peptides, are now being researched. Antimicrobial peptides, or AMPs, are peptides produced innately by the immune systems of certain organisms to fight off infection. Because they kill microorganisms through different mechanisms, they have the potential to be effective against antibiotic resistant bacteria. One such AMP is melittin, produced by honey bees and used in their venom. Melittin kills cells by forming pores in the cell membrane, eventually destabilizing the cell and causing cell lysis. Because melittin targets a conserved area of cells, bacteria cannot develop resistance to it as easily as different types of antibiotics. </p><br />
<br />
<p> Our project this summer is to transform E. coli with the melittin gene in honey bees, so that the E. coli may express the melittin peptide, thereby developing a means to mass-produce melittin as an alternative to today's antibiotics. We hope to optimize the production of melittin in E. coli, as well as to test the bioactivity of our produced melittin in comparison to its naturally-occurring counterpart. </p><br />
</div> --><br />
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Hliu1423
http://2014.igem.org/Team:Stony_Brook
Team:Stony Brook
2014-10-18T00:43:38Z
<p>Hliu1423: </p>
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background-image: url(https://static.igem.org/mediawiki/2014/a/a5/Stony_brook_home_background.png);<br />
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color: #FFF;<br />
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#facebook{<br />
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}<br />
<br />
#twitter{<br />
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padding-bottom: 5px;<br />
}<br />
<br />
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position:absolute;<br />
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margin-bottom:-107px;<br />
overflow:auto;<br />
<br />
}<br />
#projectDescription h1, p{<br />
font-family:"Trebuchet MS", Arial, Helvetica, sans-serif;<br />
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#projectDescription h1{<br />
text-align:center;<br />
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}<br />
<br />
</style><br />
<br />
</head><br />
<br />
<body><br />
<br />
<a href="https://igem.org/Main_Page"><img src="https://static.igem.org/mediawiki/2014/5/51/Stony_brook_igem_logo.png" alt= "iGEM logo" name="iGEM_logo" border="0px" id="iGEM_logo" /></a><br />
<br />
<a href="http://www.facebook.com/iGEMatstonybrook"><img src="https://static.igem.org/mediawiki/2014/0/0f/Stony_brook_fb_logo.png" alt= "facebook" name="facebook" border="0px" id="facebook" /></a></a><br />
<a href="http://twitter.com/iGEMSBU"><img src="https://static.igem.org/mediawiki/2014/b/b5/Stony_brook_twitter_logo.png" alt= "twitter" name="twitter" border="0px" id="twitter" /></a></a><br />
<br />
<br />
<div id="header"><a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/1/11/Stony_brook_header.png" alt="Stony Brook iGEM" border="0px"" /></a></div><br />
<br />
<div id="navigation_container"><br />
<br />
<div id="navigation"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/b/b8/Stony_Brook_NavIcon.Home.png" border="0px" /></a><br />
<p>Home</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Team"><img src="https://static.igem.org/mediawiki/2014/4/4a/Stony_Brook_NavIcon.Team.png" border="0px" /></a><br />
<p>Team</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Project"><img src="https://static.igem.org/mediawiki/2014/a/ab/Stony_Brook_NavIcon.Project.png" border="0px" /></a><br />
<p>Project</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Notebook"><img src="https://static.igem.org/mediawiki/2014/b/b5/Stony_Brook_NavIcon.Notebook.png" border="0px"/></a><br />
<p>Notebook</p><br />
</div><br />
<div id="navigation"><br />
<a href="http://www.igem.org"> <img src="https://static.igem.org/mediawiki/2014/c/c3/Stony_Brook_NavIcon.Results.png"/></a><br />
<p>Results</p><br />
</div><br />
<div id="navigation"><br />
<a href="http://www.igem.org"> <img src="https://static.igem.org/mediawiki/2014/e/e2/Stony_Brook_Navicon.Outreach.png" border="0px"/></a><br />
<p>Outreach</p><br />
</div> <br />
<div id="navigation"><br />
<a href="http://www.igem.org"> <img src="https://static.igem.org/mediawiki/2014/8/81/Stony_Brook_NavIcon.Safety.png"/></a><br />
<p>Safety</p><br />
</div> <br />
<div id="navigation"><br />
<a href="http://www.igem.org"> <img src="https://static.igem.org/mediawiki/2014/5/5e/Stony_Brook_NavIcon.Acknowledgements.png" border="0px"/></a><br />
<p>Attributions</p><br />
</div> <br />
</div><br />
<br />
<div id="login"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook">page</a></li><br />
<li><a href="https://2014.igem.org/Talk:Team:Stony_Brook">discussion</a></li><br />
<li><a href="https://2014.igem.org/wiki/index.php?title=Team:Stony_Brook&amp;action=edit">view source</a></li><br />
<li><a href="https://2014.igem.org/wiki/index.php?title=Team:Stony_Brook&amp;action=history">history</a></li><br />
</ul><br />
</div><br />
<br />
<!-- <div id="background"><br />
</div> --><br />
<br />
<!-- <div id="projectDescription"><br />
<h1> Project Description </h1><br />
<p> The threat of antibiotic resistance has been a concern since the discovery of penicillin. As antibiotics continue to be used to treat disease, pathogenic bacteria continue to develop resistance, eventually becoming irresponsive to multiple classes of antibiotics. This is a major problem for areas that must maintain sterile environments, such as hospitals or other health-care settings. Hospital-acquired infections (HAI) are already an existing issue that poses risks to people receiving or recovering from treatment. Gram-negative bacteria in particular are known to cause more HAIs as they are more resistant to antibiotics than gram-positive bacteria. Irresponsible use of antibiotics has only exacerbated this issue. Recent reports from the World Health Organization confirm that antibiotic resistance is no longer something to worry about in the future, but a current, global health threat. </p><br />
<br />
<p> Alternatives to modern antibiotics, such as antimicrobial peptides, are now being researched. Antimicrobial peptides, or AMPs, are peptides produced innately by the immune systems of certain organisms to fight off infection. Because they kill microorganisms through different mechanisms, they have the potential to be effective against antibiotic resistant bacteria. One such AMP is melittin, produced by honey bees and used in their venom. Melittin kills cells by forming pores in the cell membrane, eventually destabilizing the cell and causing cell lysis. Because melittin targets a conserved area of cells, bacteria cannot develop resistance to it as easily as different types of antibiotics. </p><br />
<br />
<p> Our project this summer is to transform E. coli with the melittin gene in honey bees, so that the E. coli may express the melittin peptide, thereby developing a means to mass-produce melittin as an alternative to today's antibiotics. We hope to optimize the production of melittin in E. coli, as well as to test the bioactivity of our produced melittin in comparison to its naturally-occurring counterpart. </p><br />
</div> --><br />
<br />
</body><br />
<br />
</html></div>
Hliu1423
http://2014.igem.org/Team:Stony_Brook
Team:Stony Brook
2014-10-18T00:43:04Z
<p>Hliu1423: </p>
<hr />
<div>{{CSS/Main}}<br />
<br />
<html xmlns="http://www.w3.org/1999/xhtml"><br />
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<br />
<style type="text/css"><br />
<br />
#contentSub, #footer-box, #catlinks, #search-controls, #p-logo, .printfooter, .firstHeading,.visualClear {display: none;} <br />
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background-image: url(https://static.igem.org/mediawiki/2014/a/a5/Stony_brook_home_background.png);<br />
background-repeat:no-repeat;<br />
background-position: center center;<br />
background-attachment: fixed;<br />
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-moz-background-size: cover;<br />
-o-background-size: cover;<br />
background-size: cover;<br />
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#header {<br />
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}<br />
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padding: 9px;<br />
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color: #FFF;<br />
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text-align: center;<br />
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font-weight:bold;<br />
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text-align:center;<br />
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}<br />
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a{<br />
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}<br />
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.firstHeading {<br />
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#facebook{<br />
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padding-bottom: 5px;<br />
}<br />
<br />
#twitter{<br />
position: absolute;<br />
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left: 7%;<br />
padding-bottom: 5px;<br />
}<br />
<br />
#projectDescription{<br />
position:absolute;<br />
bottom:25%;<br />
left:50%;<br />
width:584px;<br />
height:320px;<br />
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margin-bottom:-107px;<br />
overflow:auto;<br />
<br />
}<br />
#projectDescription h1, p{<br />
font-family:"Trebuchet MS", Arial, Helvetica, sans-serif;<br />
color:#FFFFFF;<br />
padding: 0px 20px 0px 20px;<br />
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}<br />
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#projectDescription h1{<br />
text-align:center;<br />
}<br />
<br />
#projectDescription p{<br />
text-indent: 25px;<br />
}<br />
<br />
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position:absolute;<br />
bottom:25%;<br />
left:50%;<br />
width:584px;<br />
height:320px;<br />
background-color:#000;<br />
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filter:alpha(opacity=60);<br />
}<br />
<br />
</style><br />
<br />
</head><br />
<br />
<body><br />
<br />
<a href="https://igem.org/Main_Page"><img src="https://static.igem.org/mediawiki/2014/5/51/Stony_brook_igem_logo.png" alt= "iGEM logo" name="iGEM_logo" border="0px" id="iGEM_logo" /></a><br />
<br />
<a href="http://www.facebook.com/iGEMatstonybrook"><img src="https://static.igem.org/mediawiki/2014/0/0f/Stony_brook_fb_logo.png" alt= "facebook" name="facebook" border="0px" id="facebook" /></a></a><br />
<a href="http://twitter.com/iGEMSBU"><img src="https://static.igem.org/mediawiki/2014/b/b5/Stony_brook_twitter_logo.png" alt= "twitter" name="twitter" border="0px" id="twitter" /></a></a><br />
<br />
<br />
<div id="header"><a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/1/11/Stony_brook_header.png" alt="Stony Brook iGEM" border="0px"" /></a></div><br />
<br />
<div id="navigation_container"><br />
<br />
<div id="navigation"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/b/b8/Stony_Brook_NavIcon.Home.png" border="0px" /></a><br />
<p>Home</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Team"><img src="https://static.igem.org/mediawiki/2014/4/4a/Stony_Brook_NavIcon.Team.png" border="0px" /></a><br />
<p>Team</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Project"><img src="https://static.igem.org/mediawiki/2014/a/ab/Stony_Brook_NavIcon.Project.png" border="0px" /></a><br />
<p>Project</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Notebook"><img src="https://static.igem.org/mediawiki/2014/b/b5/Stony_Brook_NavIcon.Notebook.png" border="0px"/></a><br />
<p>Notebook</p><br />
</div><br />
<div id="navigation"><br />
<a href="http://www.igem.org"> <img src="https://static.igem.org/mediawiki/2014/c/c3/Stony_Brook_NavIcon.Results.png"/></a><br />
<p>Results</p><br />
</div><br />
<div id="navigation"><br />
<a href="http://www.igem.org"> <img src="https://static.igem.org/mediawiki/2014/e/e2/Stony_Brook_Navicon.Outreach.png" border="0px"/></a><br />
<p>Outreach</p><br />
</div> <br />
<div id="navigation"><br />
<a href="http://www.igem.org"> <img src="https://static.igem.org/mediawiki/2014/8/81/Stony_Brook_NavIcon.Safety.png"/></a><br />
<p>Safety</p><br />
</div> <br />
<div id="navigation"><br />
<a href="http://www.igem.org"> <img src="https://static.igem.org/mediawiki/2014/5/5e/Stony_Brook_NavIcon.Acknowledgements.png" border="0px"/></a><br />
<p>Attributions</p><br />
</div> <br />
</div><br />
<br />
<div id="login"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook">page</a></li><br />
<li><a href="https://2014.igem.org/Talk:Team:Stony_Brook">discussion</a></li><br />
<li><a href="https://2014.igem.org/wiki/index.php?title=Team:Stony_Brook&amp;action=edit">view source</a></li><br />
<li><a href="https://2014.igem.org/wiki/index.php?title=Team:Stony_Brook&amp;action=history">history</a></li><br />
</ul><br />
</div><br />
<br />
<!-- <div id="background"><br />
</div> --><br />
<br />
<!-- <div id="projectDescription"><br />
<h1> Project Description </h1><br />
<p> The threat of antibiotic resistance has been a concern since the discovery of penicillin. As antibiotics continue to be used to treat disease, pathogenic bacteria continue to develop resistance, eventually becoming irresponsive to multiple classes of antibiotics. This is a major problem for areas that must maintain sterile environments, such as hospitals or other health-care settings. Hospital-acquired infections (HAI) are already an existing issue that poses risks to people receiving or recovering from treatment. Gram-negative bacteria in particular are known to cause more HAIs as they are more resistant to antibiotics than gram-positive bacteria. Irresponsible use of antibiotics has only exacerbated this issue. Recent reports from the World Health Organization confirm that antibiotic resistance is no longer something to worry about in the future, but a current, global health threat. </p><br />
<br />
<p> Alternatives to modern antibiotics, such as antimicrobial peptides, are now being researched. Antimicrobial peptides, or AMPs, are peptides produced innately by the immune systems of certain organisms to fight off infection. Because they kill microorganisms through different mechanisms, they have the potential to be effective against antibiotic resistant bacteria. One such AMP is melittin, produced by honey bees and used in their venom. Melittin kills cells by forming pores in the cell membrane, eventually destabilizing the cell and causing cell lysis. Because melittin targets a conserved area of cells, bacteria cannot develop resistance to it as easily as different types of antibiotics. </p><br />
<br />
<p> Our project this summer is to transform E. coli with the melittin gene in honey bees, so that the E. coli may express the melittin peptide, thereby developing a means to mass-produce melittin as an alternative to today's antibiotics. We hope to optimize the production of melittin in E. coli, as well as to test the bioactivity of our produced melittin in comparison to its naturally-occurring counterpart. </p><br />
</div> --><br />
<br />
</body><br />
<br />
</html></div>
Hliu1423
http://2014.igem.org/Team:Stony_Brook
Team:Stony Brook
2014-10-18T00:40:09Z
<p>Hliu1423: </p>
<hr />
<div>{{CSS/Main}}<br />
<br />
<html xmlns="http://www.w3.org/1999/xhtml"><br />
<br />
<br />
<style type="text/css"><br />
<br />
#contentSub, #footer-box, #catlinks, #search-controls, #p-logo, .printfooter, .firstHeading,.visualClear {display: none;} <br />
<br />
body {<br />
background-image: url(https://static.igem.org/mediawiki/2014/a/a5/Stony_brook_home_background.png);<br />
background-repeat:no-repeat;<br />
background-position: center center;<br />
background-attachment: fixed;<br />
-webkit-background-size: cover;<br />
-moz-background-size: cover;<br />
-o-background-size: cover;<br />
background-size: cover;<br />
}<br />
#header {<br />
width: 672px;<br />
position: absolute;<br />
top:29%;<br />
left:50%;<br />
margin-left: -310px;<br />
margin-top:-50px;<br />
}<br />
<br />
#navigation {<br />
width: 65px;<br />
padding: 9px;<br />
float: left;<br />
color: #FFF;<br />
font-family:"Trebuchet MS", Arial, Helvetica, sans-serif;<br />
text-align: center;<br />
font-size:small;<br />
font-weight:bold;<br />
}<br />
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#navigation p{<br />
display:none;<br />
text-align:center;<br />
}<br />
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#navigation a:hover + p{<br />
display: block;<br />
}<br />
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img:hover{<br />
opacity: 0.8;<br />
filter: alpha(opacity=80);<br />
}<br />
<br />
#iGEM_logo {<br />
position: absolute;<br />
left: 90%;<br />
top: 2%;<br />
}<br />
<br />
#navigation_container {<br />
width: 800 px;<br />
position: absolute;<br />
top: 40%;<br />
right: 50%;<br />
margin-right: -355px;<br />
}<br />
<br />
<br />
<br />
a{<br />
outline: 0;<br />
}<br />
<br />
#login{<br />
width: 400px;<br />
height: 20px;<br />
background-color: #000;<br />
opacity: 0.7;<br />
filter: alpha(opacity=70);<br />
position: absolute;<br />
top: 0;<br />
left:0;<br />
}<br />
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list-style-type: none;<br />
margin: 0;<br />
padding: 0px;<br />
}<br />
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#login li{<br />
float: left;<br />
text-align: center;<br />
}<br />
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#login a{<br />
display: block;<br />
height: 20px;<br />
line-height: 20px;<br />
width: 100px;<br />
text-decoration: none;<br />
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font-size: x-small;<br />
vertical-align: middle;<br />
font-weight: bold;<br />
color: #FFF;<br />
}<br />
<br />
#login a:hover{<br />
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}<br />
<br />
.firstHeading {<br />
visibility: hidden;<br />
}<br />
<br />
#facebook{<br />
position: absolute;<br />
bottom: 1%;<br />
left: 1%;<br />
padding-bottom: 5px;<br />
}<br />
<br />
#twitter{<br />
position: absolute;<br />
bottom: 1%;<br />
left: 7%;<br />
padding-bottom: 5px;<br />
}<br />
<br />
#projectDescription{<br />
position:absolute;<br />
bottom:25%;<br />
left:50%;<br />
width:584px;<br />
height:320px;<br />
margin-left:-265.6px;<br />
margin-bottom:-107px;<br />
overflow:auto;<br />
<br />
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font-family:"Trebuchet MS", Arial, Helvetica, sans-serif;<br />
color:#FFFFFF;<br />
padding: 0px 20px 0px 20px;<br />
opacity:1;<br />
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#projectDescription h1{<br />
text-align:center;<br />
}<br />
<br />
#projectDescription p{<br />
text-indent: 25px;<br />
}<br />
<br />
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bottom:25%;<br />
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width:584px;<br />
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background-color:#000;<br />
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margin-bottom:-107px;<br />
color: #FFF;<br />
opacity:0.6;<br />
filter:alpha(opacity=60);<br />
}<br />
<br />
</style><br />
<br />
</head><br />
<br />
<body><br />
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<!-- <div id="projectDescription"><br />
<h1> Project Description </h1><br />
<p> The threat of antibiotic resistance has been a concern since the discovery of penicillin. As antibiotics continue to be used to treat disease, pathogenic bacteria continue to develop resistance, eventually becoming irresponsive to multiple classes of antibiotics. This is a major problem for areas that must maintain sterile environments, such as hospitals or other health-care settings. Hospital-acquired infections (HAI) are already an existing issue that poses risks to people receiving or recovering from treatment. Gram-negative bacteria in particular are known to cause more HAIs as they are more resistant to antibiotics than gram-positive bacteria. Irresponsible use of antibiotics has only exacerbated this issue. Recent reports from the World Health Organization confirm that antibiotic resistance is no longer something to worry about in the future, but a current, global health threat. </p><br />
<br />
<p> Alternatives to modern antibiotics, such as antimicrobial peptides, are now being researched. Antimicrobial peptides, or AMPs, are peptides produced innately by the immune systems of certain organisms to fight off infection. Because they kill microorganisms through different mechanisms, they have the potential to be effective against antibiotic resistant bacteria. One such AMP is melittin, produced by honey bees and used in their venom. Melittin kills cells by forming pores in the cell membrane, eventually destabilizing the cell and causing cell lysis. Because melittin targets a conserved area of cells, bacteria cannot develop resistance to it as easily as different types of antibiotics. </p><br />
<br />
<p> Our project this summer is to transform E. coli with the melittin gene in honey bees, so that the E. coli may express the melittin peptide, thereby developing a means to mass-produce melittin as an alternative to today's antibiotics. We hope to optimize the production of melittin in E. coli, as well as to test the bioactivity of our produced melittin in comparison to its naturally-occurring counterpart. </p><br />
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Hliu1423
http://2014.igem.org/Team:Stony_Brook/Notebook
Team:Stony Brook/Notebook
2014-10-17T23:37:06Z
<p>Hliu1423: </p>
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<div id="yellow_header"> <a href="http://www.stonybrook.edu/">Stony Brook University</a></div><br />
<div id="blue_header"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/1/1e/Stony_Brook_Apis-biotics_header.png"/ id="apis-biotics"/></a><br />
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<p> Home </p><br />
</div><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook/Team"><img src="https://static.igem.org/mediawiki/2014/6/66/Stony_Brook_NavIcon2_Team.png"/></a><br />
<p> Team </p><br />
</div><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook/Project"><img src="https://static.igem.org/mediawiki/2014/9/99/Stony_Brook_NavIcon2_Project.png"/></a><br />
<p> Project </p><br />
</div><br />
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<p> Notebook </p><br />
</div><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/8/80/Stony_Brook_NavIcon2_Outreach.png"/></a><br />
<p> Outreach </p><br />
</div><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/0/03/Stony_Brook_NavIcon2_Acknowledgements.png"/></a><br />
<p> Attributions </p><br />
</div><br />
</div><br />
</div><br />
<div id="project"><br />
<p>Notebook</p><br />
</div><br />
<br />
<div class="picture"><img src="https://static.igem.org/mediawiki/2014/0/00/Stony_Brook_LabSpace.png" /></div><br />
<br />
</div><br />
<br />
<p class="glossary"><a href="https://2014.igem.org/Team:Stony_Brook/MaterialsMethods">Click here to see our procedures</a></p><br />
<br />
<div id="contents"> <section class="ac-container"><br />
<section class="ac-container"><br />
<div><br />
<input id="ac-1" name="accordion-1" type="checkbox" /><br />
<label for="ac-1">June</label><br />
<article class="ac-large"><br />
<br/><br/><h4>Week 1</h4><br />
<p>Literary research: </p><br />
<p>We looked into SUMO protein, a protective group that we could use to stabilize the melittin peptide. We also looked into mechanisms to express or release melittin:<br /><br />
<ul><li> Using a signalling sequence, signal peptides and pathways (OmpA, SecB, IgA beta)in order to secrete melittin</li><br />
<li>Lysing our cells with a killswitch and then using FLAG tags, affinity tags to purify melittin from lysate</li><br />
<li>Proteases to cleave off a protecting group.</li></ul></p> <br />
<p> We also looked into understanding biobricking assembly standards</p><br />
<br />
<h4>Week 2</h4> <br />
<p> Literary research:</p><br />
<ul> <li>Prepromelittin nucleotide sequence was found.</li><br />
<li> Decided on a fusion protein with melittin and GST tag</li><br />
<li>Reached out to Genomics Core Facility on Stony Brook University Campus to talk with Dr. John Schwedes about RNA extraction and Dr. Dmitri Gnatenko about primer design.</li><br />
<li>Searching for plasmids: met with Yelena from Molecular Cloning Facility to discuss appropriate plasmids for our experiment and search through their plasmid library</li></ul><br />
<p>Experimental:</p><br />
<ul><li>Competent E.coli cell line made. </li><br />
<li> Visited local beehive to obtain Apis mellifera samples; dissected our bees and extracted their RNA </li><br />
<li>Transformed RhlR biobrick</li></ul><br />
<p> Outreach:</p><br />
<p>We presented on synthetic biology at four classes at a local high school, East Sachem High School!</p><br />
<br />
<h4> Week 3</h4><br />
<p>Literary research:</p><br />
<ul><li> Looking into P. syringae as a model for P. aeruginosa?</li><br />
<li> Optimal expression conditions</li><br />
<li> Looking into TEV protease site to cleave off GST protein</li><br />
<li>Detection methods</li></ul><br />
<p>Experimental:</p><br />
<ul><li> Designed primers for melittin amplification and tagged melittin</li><br />
<li>Successful RNA extraction!</li><br />
<li>Obtained plasmid from our advisor, Dr. Czaplinski</li><br />
<li>Designed RhlR circuit and obtained signaling compound C4HSL</li><br />
<li>Constitutive promoter BBa_J23101 and J23102 transformed for later fluorescence testing</li></ul><br />
<br />
<h4>Week 4</h4><br />
<p>Experimental:</p><br />
<ul><li>Synthesized cDNA library</li><br />
<li> Designed and ordered Biobrick melittin primers</li><br />
<li>mCherry and fluorescence testing</li></ul><br />
<p>Outreach:</p><br />
<ul><li>Looked into doing an iGEM survey with other schools with the Stony Brook Institutional Review Board </li><br />
<li> Attended iGEM High School Jamboree!</li></ul><br />
</article><br />
</div><br />
<div><br />
<input id="ac-2" name="accordion-1" type="checkbox"><br />
<label for="ac-2">July</label><br />
<article class="ac-largeish"><br />
<br/><br/><h4>Week 1</h4><br />
<p>Experimental:</p><br />
<ul><li> Ligated and transformed for GST-melittin fusion protein</li><br />
<li>Induced plasmid with IPTG and checked with SDS page gel- no protein detected </li><br />
<li>Redid PCR and digest</li><br />
<li>Sequenced RhlR/Rhl circuit samples</li></ul><br />
<br />
<h4>Week 2</h4><br />
<p>Experimental:</p><br />
<ul><li>Prepared insert for our melittin plasmid- used PCR to amplify melittin, insert TEV cleavage site, then digested insert, and purified</li><br />
<li>Digested RhlR/ rhl plasmid, phosphatased, purified, ligated, and transformed. Cultured overnight, miniprepped and ran on gel to check results.</li><br />
<li>RhlR + Promoter miniprep, sent for sequencing</li></ul><br />
<p>Outreach</p><br />
<p>Presentated at Stony Brook University Biotechnology Camp for high school students!</p><br />
<h4>Week 3</h4><br />
<p>Experimental:</p><br />
<ul><li>Digest check, sequencing results for RhlR/Rhl</li><br />
<li>Screened colonies for melittin </li><br />
<li>Began PCR from beginning to amplify prepromelittin</li><br />
add-on PCR, inserted into duet plasmid</li></ul><br />
<p> Mathematical modeling:</p><br />
<p>Researching articles for rates of RhlR degradation</p><br />
<p>Comparison of our model with arbitrary values to experimental model for fluorescence</p><br />
<br />
<br />
<h4>Week 4</h4><br />
<p>Experimental:</p><br />
<ul><li>PCR amplification of prepromelittin from cDNA, addition of TEV site to melittin gene</li><br />
<li>Ligation of TEV-melittin into GST plasmid</li><br />
<li>Transformation of E.coli with ligation</li><br />
<li>Checked successful transformation by digesting transformed plasmid and viewing diegest on gel to cross-check with expected band size-70% success! </li><br />
<li>Samples from transformations sent for senquencing</li><br />
<li>N17+RhlR ligation into duet plasmid troubleshooting</li></ul><br />
<p> Mathematical modeling:</p><br />
<ul><li>Rates for C4HSL and RhlR degradation, dimerization</li><br />
<li>Preliminary mathematical models created!</li></ul><br />
<br />
</article><br />
</div><br />
<div><br />
<input id="ac-3" name="accordion-1" type="checkbox"><br />
<label for="ac-3">August</label><br />
<article class="ac-small"><br />
<br/><br/> <h4>Week 1</h4><br />
<p>Experimental:</p><br />
<ul><li>Results from sequencing: 3/4 samples were in frame; 1 had a missene mutation</li><br />
<li>Inducing of cultures of GST plasmid (control) and GST+TEV+melittin with IPTG</li><br />
<li> Attempted to checked protein expression by SDS-page</li><br />
<li>Co-transformations of RhlR plasmids</li></ul><br />
<h4>Week 2</h4><br />
<ul><li>Expression of melittin confirmed by SDS-PAGE!</li><br />
<li>Completion of Rhl circuit</li><br />
<li>Samples sent in for sequencing</li><br />
</ul><br />
<h4>Week 3</h4><br />
<p>School starts! </p><br />
</article><br />
</div><br />
<div><br />
<input id="ac-4" name="accordion-1" type="checkbox"><br />
<label for="ac-4">September</label><br />
<article class="ac-med"><br />
<br/><br/> <h4>Week 1:</h4><br />
<p>Experimental:</p><br />
<ul><li>Expression of melittin</li><br />
<li>Checked protein expression by SDS-PAGE</li><br />
<li>Ligation of Rhl into mCherry plasmid</li><br />
<li> Double digestion to verify that ligation was successful</li></ul><br />
<p> Mathematical Modeling</p><br />
<ul><li>Adjusting model for protein expression</li><br />
<li>Visualization of melittin expression using MATLAB</li></ul><br />
<p>Outreach:</p><br />
<ul><li>Feature in Stony Brook Scholars Newsletter</li><br />
<li>Stony Brook involvement fair!</li></ul><br />
<br />
<h4>Week 2:</h4><br />
<p>Experimental:</p><ul><br />
<li>Measuring of fluroresence expression using TECAN </li><br />
<li>Found that Rhl is a leaky promoter, performed experiments to validate our concerns</li></ul><br />
<br />
<h4>Week 3</h4><br />
<ul><li>Culturing of cells with GST+TEV+Melittin</li><br />
<li>Checking protein expression by SDS-PAGE</li><br />
<li> Biobricking</li></ul><br />
</article><br />
</div><br />
</section> <br />
</div><br />
</body><br />
</html></div>
Hliu1423
http://2014.igem.org/Team:Stony_Brook/Notebook
Team:Stony Brook/Notebook
2014-10-17T23:36:35Z
<p>Hliu1423: </p>
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<div id="yellow_header"> <a href="http://www.stonybrook.edu/">Stony Brook University</a></div><br />
<div id="blue_header"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/1/1e/Stony_Brook_Apis-biotics_header.png"/ id="apis-biotics"/></a><br />
<div id="yellow_navigation_container"><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/7/78/Stony_Brook_NavIcon2_Home.png"/></a><br />
<p> Home </p><br />
</div><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook/Team"><img src="https://static.igem.org/mediawiki/2014/6/66/Stony_Brook_NavIcon2_Team.png"/></a><br />
<p> Team </p><br />
</div><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook/Project"><img src="https://static.igem.org/mediawiki/2014/9/99/Stony_Brook_NavIcon2_Project.png"/></a><br />
<p> Project </p><br />
</div><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook/Notebook"><img src="https://static.igem.org/mediawiki/2014/5/57/Stony_Brook_NavIcon2_Notebook.png"/></a><br />
<p> Notebook </p><br />
</div><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/8/80/Stony_Brook_NavIcon2_Outreach.png"/></a><br />
<p> Outreach </p><br />
</div><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/0/03/Stony_Brook_NavIcon2_Acknowledgements.png"/></a><br />
<p> Attributions </p><br />
</div><br />
</div><br />
</div><br />
<div id="project"><br />
<p>Notebook</p><br />
</div><br />
<br />
<div class="picture"><img src="https://static.igem.org/mediawiki/2014/0/00/Stony_Brook_LabSpace.png" /></div><br />
<br />
</div><br />
<br />
<p class="glossary"><a href="https://2014.igem.org/Team:Stony_Brook/MaterialsMethods">Click here to see our procedures</a></p><br />
<br />
<div id="contents"> <section class="ac-container"><br />
<section class="ac-container"><br />
<div><br />
<input id="ac-1" name="accordion-1" type="checkbox" /><br />
<label for="ac-1">June</label><br />
<article class="ac-large"><br />
<br/><br/><h4>Week 1</h4><br />
<p>Literary research: </p><br />
<p>We looked into SUMO protein, a protective group that we could use to stabilize the melittin peptide. We also looked into mechanisms to express or release melittin:<br /><br />
<ul><li> Using a signalling sequence, signal peptides and pathways (OmpA, SecB, IgA beta)in order to secrete melittin</li><br />
<li>Lysing our cells with a killswitch and then using FLAG tags, affinity tags to purify melittin from lysate</li><br />
<li>Proteases to cleave off a protecting group.</li></ul></p> <br />
<p> We also looked into understanding biobricking assembly standards</p><br />
<br />
<h4>Week 2</h4> <br />
<p> Literary research:</p><br />
<ul> <li>Prepromelittin nucleotide sequence was found.</li><br />
<li> Decided on a fusion protein with melittin and GST tag</li><br />
<li>Reached out to Genomics Core Facility on Stony Brook University Campus to talk with Dr. John Schwedes about RNA extraction and Dr. Dmitri Gnatenko about primer design.</li><br />
<li>Searching for plasmids: met with Yelena from Molecular Cloning Facility to discuss appropriate plasmids for our experiment and search through their plasmid library</li></ul><br />
<p>Experimental:</p><br />
<ul><li>Competent E.coli cell line made. </li><br />
<li> Visited local beehive to obtain Apis mellifera samples; dissected our bees and extracted their RNA </li><br />
<li>Transformed RhlR biobrick</li></ul><br />
<p> Outreach:</p><br />
<p>We presented on synthetic biology at four classes at a local high school, East Sachem High School!</p><br />
<br />
<h4> Week 3</h4><br />
<p>Literary research:</p><br />
<ul><li> Looking into P. syringae as a model for P. aeruginosa?</li><br />
<li> Optimal expression conditions</li><br />
<li> Looking into TEV protease site to cleave off GST protein</li><br />
<li>Detection methods</li></ul><br />
<p>Experimental:</p><br />
<ul><li> Designed primers for melittin amplification and tagged melittin</li><br />
<li>Successful RNA extraction! (Picture from Week 7/2)</li><br />
<li>Obtained plasmid from our advisor, Dr. Czaplinski</li><br />
<li>Designed RhlR circuit and obtained signaling compound C4HSL</li><br />
<li>Constitutive promoter BBa_J23101 and J23102 transformed for later fluorescence testing</li></ul><br />
<br />
<h4>Week 4</h4><br />
<p>Experimental:</p><br />
<ul><li>Synthesized cDNA library</li><br />
<li> Designed and ordered Biobrick melittin primers</li><br />
<li>mCherry and fluorescence testing</li></ul><br />
<p>Outreach:</p><br />
<ul><li>Looked into doing an iGEM survey with other schools with the Stony Brook Institutional Review Board </li><br />
<li> Attended iGEM High School Jamboree! (picture) </li></ul><br />
</article><br />
</div><br />
<div><br />
<input id="ac-2" name="accordion-1" type="checkbox"><br />
<label for="ac-2">July</label><br />
<article class="ac-largeish"><br />
<br/><br/><h4>Week 1</h4><br />
<p>Experimental:</p><br />
<ul><li> Ligated and transformed for GST-melittin fusion protein</li><br />
<li>Induced plasmid with IPTG and checked with SDS page gel- no protein detected </li><br />
<li>Redid PCR and digest</li><br />
<li>Sequenced RhlR/Rhl circuit samples</li></ul><br />
<br />
<h4>Week 2</h4><br />
<p>Experimental:</p><br />
<ul><li>Prepared insert for our melittin plasmid- used PCR to amplify melittin, insert TEV cleavage site, then digested insert, and purified</li><br />
<li>Digested RhlR/ rhl plasmid, phosphatased, purified, ligated, and transformed. Cultured overnight, miniprepped and ran on gel to check results.</li><br />
<li>RhlR + Promoter miniprep, sent for sequencing</li></ul><br />
<p>Outreach</p><br />
<p>Presentated at Stony Brook University Biotechnology Camp for high school students!</p><br />
<h4>Week 3</h4><br />
<p>Experimental:</p><br />
<ul><li>Digest check, sequencing results for RhlR/Rhl</li><br />
<li>Screened colonies for melittin </li><br />
<li>Began PCR from beginning to amplify prepromelittin</li><br />
add-on PCR, inserted into duet plasmid</li></ul><br />
<p> Mathematical modeling:</p><br />
<p>Researching articles for rates of RhlR degradation</p><br />
<p>Comparison of our model with arbitrary values to experimental model for fluorescence</p><br />
<br />
<br />
<h4>Week 4</h4><br />
<p>Experimental:</p><br />
<ul><li>PCR amplification of prepromelittin from cDNA, addition of TEV site to melittin gene</li><br />
<li>Ligation of TEV-melittin into GST plasmid</li><br />
<li>Transformation of E.coli with ligation</li><br />
<li>Checked successful transformation by digesting transformed plasmid and viewing diegest on gel to cross-check with expected band size-70% success! </li><br />
<li>Samples from transformations sent for senquencing</li><br />
<li>N17+RhlR ligation into duet plasmid troubleshooting</li></ul><br />
<p> Mathematical modeling:</p><br />
<ul><li>Rates for C4HSL and RhlR degradation, dimerization</li><br />
<li>Preliminary mathematical models created!</li></ul><br />
<br />
</article><br />
</div><br />
<div><br />
<input id="ac-3" name="accordion-1" type="checkbox"><br />
<label for="ac-3">August</label><br />
<article class="ac-small"><br />
<br/><br/> <h4>Week 1</h4><br />
<p>Experimental:</p><br />
<ul><li>Results from sequencing: 3/4 samples were in frame; 1 had a missene mutation</li><br />
<li>Inducing of cultures of GST plasmid (control) and GST+TEV+melittin with IPTG</li><br />
<li> Attempted to checked protein expression by SDS-page</li><br />
<li>Co-transformations of RhlR plasmids</li></ul><br />
<h4>Week 2</h4><br />
<ul><li>Expression of melittin confirmed by SDS-PAGE!</li><br />
<li>Completion of Rhl circuit</li><br />
<li>Samples sent in for sequencing</li><br />
</ul><br />
<h4>Week 3</h4><br />
<p>School starts! </p><br />
</article><br />
</div><br />
<div><br />
<input id="ac-4" name="accordion-1" type="checkbox"><br />
<label for="ac-4">September</label><br />
<article class="ac-med"><br />
<br/><br/> <h4>Week 1:</h4><br />
<p>Experimental:</p><br />
<ul><li>Expression of melittin</li><br />
<li>Checked protein expression by SDS-PAGE</li><br />
<li>Ligation of Rhl into mCherry plasmid</li><br />
<li> Double digestion to verify that ligation was successful</li></ul><br />
<p> Mathematical Modeling</p><br />
<ul><li>Adjusting model for protein expression</li><br />
<li>Visualization of melittin expression using MATLAB</li></ul><br />
<p>Outreach:</p><br />
<ul><li>Feature in Stony Brook Scholars Newsletter</li><br />
<li>Stony Brook involvement fair!</li></ul><br />
<br />
<h4>Week 2:</h4><br />
<p>Experimental:</p><ul><br />
<li>Measuring of fluroresence expression using TECAN </li><br />
<li>Found that Rhl is a leaky promoter, performed experiments to validate our concerns</li></ul><br />
<br />
<h4>Week 3</h4><br />
<ul><li>Culturing of cells with GST+TEV+Melittin</li><br />
<li>Checking protein expression by SDS-PAGE</li><br />
<li> Biobricking</li></ul><br />
</article><br />
</div><br />
</section> <br />
</div><br />
</body><br />
</html></div>
Hliu1423
http://2014.igem.org/Team:Stony_Brook/Notebook
Team:Stony Brook/Notebook
2014-10-17T23:35:59Z
<p>Hliu1423: </p>
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<div id="yellow_header"> <a href="http://www.stonybrook.edu/">Stony Brook University</a></div><br />
<div id="blue_header"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/1/1e/Stony_Brook_Apis-biotics_header.png"/ id="apis-biotics"/></a><br />
<div id="yellow_navigation_container"><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/7/78/Stony_Brook_NavIcon2_Home.png"/></a><br />
<p> Home </p><br />
</div><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook/Team"><img src="https://static.igem.org/mediawiki/2014/6/66/Stony_Brook_NavIcon2_Team.png"/></a><br />
<p> Team </p><br />
</div><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook/Project"><img src="https://static.igem.org/mediawiki/2014/9/99/Stony_Brook_NavIcon2_Project.png"/></a><br />
<p> Project </p><br />
</div><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook/Notebook"><img src="https://static.igem.org/mediawiki/2014/5/57/Stony_Brook_NavIcon2_Notebook.png"/></a><br />
<p> Notebook </p><br />
</div><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/8/80/Stony_Brook_NavIcon2_Outreach.png"/></a><br />
<p> Outreach </p><br />
</div><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/0/03/Stony_Brook_NavIcon2_Acknowledgements.png"/></a><br />
<p> Attributions </p><br />
</div><br />
</div><br />
</div><br />
<div id="project"><br />
<p>Notebook</p><br />
</div><br />
<br />
<div class="picture"><img src="https://static.igem.org/mediawiki/2014/0/00/Stony_Brook_LabSpace.png" /></div><br />
<br />
</div><br />
<br />
<p class="glossary"><a href="https://2014.igem.org/Team:Stony_Brook/MaterialsMethods">Click here to see our procedures</a></p><br />
<br />
<div id="contents"> <section class="ac-container"><br />
<section class="ac-container"><br />
<div><br />
<input id="ac-1" name="accordion-1" type="checkbox" /><br />
<label for="ac-1">June</label><br />
<article class="ac-large"><br />
<br/><br/><h4>Week 1</h4><br />
<p>Literary research: </p><br />
<p>We looked into SUMO protein, a protective group that we could use to stabilize the melittin peptide. We also looked into mechanisms to express or release melittin:<br /><br />
<ul><li> Using a signalling sequence, signal peptides and pathways (OmpA, SecB, IgA beta)in order to secrete melittin</li><br />
<li>Lysing our cells with a killswitch and then using FLAG tags, affinity tags to purify melittin from lysate</li><br />
<li>Proteases to cleave off a protecting group.</li></ul></p> <br />
<p> We also looked into understanding biobricking assembly standards</p><br />
<br />
<h4>Week 2</h4> <br />
<p> Literary research:</p><br />
<ul> <li>Prepromelittin nucleotide sequence was found.</li><br />
<li> Decided on a fusion protein with melittin and GST tag</li><br />
<li>Reached out to Genomics Core Facility on Stony Brook University Campus to talk with Dr. John Schwedes about RNA extraction and Dr. Dmitri Gnatenko about primer design.</li><br />
<li>Searching for plasmids: met with Yelena from Molecular Cloning Facility to discuss appropriate plasmids for our experiment and search through their plasmid library</li></ul><br />
<p>Experimental:</p><br />
<ul><li>Competent E.coli cell line made. </li><br />
<li> Visited local beehive to obtain Apis mellifera samples; dissected our bees and extracted their RNA </li><br />
<li>Transformed RhlR biobrick</li></ul><br />
<p> Outreach:</p><br />
<p>We presented on synthetic biology at four classes at a local high school, East Sachem High School!</p><br />
<br />
<h4> Week 3</h4><br />
<p>Literary research:</p><br />
<ul><li> Looking into P. syringae as a model for P. aeruginosa?</li><br />
<li> Optimal expression conditions</li><br />
<li> Looking into TEV protease site to cleave off GST protein</li><br />
<li>Detection methods</li></ul><br />
<p>Experimental:</p><br />
<ul><li> Designed primers for melittin amplification and tagged melittin</li><br />
<li>Successful RNA extraction! (Picture from Week 7/2)</li><br />
<li>Obtained plasmid from our advisor, Dr. Czaplinski</li><br />
<li>Designed RhlR circuit and obtained signaling compound C4HSL</li><br />
<li>Constitutive promoter BBa_J23101 and J23102 transformed for later fluorescence testing</li></ul><br />
<br />
<h4>Week 4</h4><br />
<p>Experimental:</p><br />
<ul><li>Synthesized cDNA library</li><br />
<li> Designed and ordered Biobrick melittin primers</li><br />
<li>mCherry and fluorescence testing</li></ul><br />
<p>Outreach:</p><br />
<ul><li>Looked into doing an iGEM survey with other schools with the Stony Brook Institutional Review Board </li><br />
<li> Attended iGEM High School Jamboree! (picture) </li></ul><br />
</article><br />
</div><br />
<div><br />
<input id="ac-2" name="accordion-1" type="checkbox"><br />
<label for="ac-2">July</label><br />
<article class="ac-largeish"><br />
<br/><br/><h4>Week 1</h4><br />
<p>Experimental:</p><br />
<ul><li> Ligated and transformed for GST-melittin fusion protein</li><br />
<li>Induced plasmid with IPTG and checked with SDS page gel- no protein detected </li><br />
<li>Redid PCR and digest</li><br />
<li>Sequenced RhlR/Rhl circuit samples</li></ul><br />
<br />
<h4>Week 2</h4><br />
<p>Experimental:</p><br />
<ul><li>Prepared insert for our melittin plasmid- used PCR to amplify melittin, insert TEV cleavage site, then digested insert, and purified</li><br />
<li>Digested RhlR/ rhl plasmid, phosphatased, purified, ligated, and transformed. Cultured overnight, miniprepped and ran on gel to check results.</li><br />
<li>RhlR + Promoter miniprep, sent for sequencing</li></ul><br />
<p>Outreach</p><br />
<p>Presentated at Stony Brook University Biotechnology Camp for high school students!</p><br />
<h4>Week 3</h4><br />
<p>Experimental:</p><br />
<ul><li>Digest check, sequencing results for RhlR/Rhl</li><br />
<li>Screened colonies for melittin </li><br />
<li>Began PCR from beginning to amplify prepromelittin</li><br />
add-on PCR, inserted into duet plasmid</li></ul><br />
<p> Mathematical modeling:</p><br />
<p>Researching articles for rates of RhlR degradation</p><br />
<p>Comparison of our model with arbitrary values to experimental model for fluorescence</p><br />
<br />
<br />
<h4>Week 4</h4><br />
<p>Experimental:</p><br />
<ul><li>PCR amplification of prepromelittin from cDNA, addition of TEV site to melittin gene</li><br />
<li>Ligation of TEV-melittin into GST plasmid</li><br />
<li>Transformation of E.coli with ligation</li><br />
<li>Checked successful transformation by digesting transformed plasmid and viewing diegest on gel to cross-check with expected band size-70% success! </li><br />
<li>Samples from transformations sent for senquencing</li><br />
<li>N17+RhlR ligation into duet plasmid troubleshooting</li></ul><br />
<p> Mathematical modeling:</p><br />
<ul><li>Rates for C4HSL and RhlR degradation, dimerization</li><br />
<li>Preliminary mathematical models created!</li></ul><br />
<br />
</article><br />
</div><br />
<div><br />
<input id="ac-3" name="accordion-1" type="checkbox"><br />
<label for="ac-3">August</label><br />
<article class="ac-small"><br />
<br/><br/> <h4>Week 1</h4><br />
<p>Experimental:</p><br />
<ul><li>Results from sequencing: 3/4 samples were in frame; 1 had a missene mutation</li><br />
<li>Inducing of cultures of GST plasmid (control) and GST+TEV+melittin with IPTG</li><br />
<li> Attempted to checked protein expression by SDS-page</li><br />
<li>Co-transformations of RhlR plasmids</li></ul><br />
<h4>Week 2</h4><br />
<ul><li>Expression of melittin confirmed by SDS-PAGE!</li><br />
<li>Completion of Rhl circuit</li><br />
<li>Samples sent in for sequencing</li><br />
</ul><br />
<h4>Week 3</h4><br />
<p>School starts! </p><br />
</article><br />
</div><br />
<div><br />
<input id="ac-4" name="accordion-1" type="checkbox"><br />
<label for="ac-4">September</label><br />
<article class="ac-med"><br />
<br/><br/> <h4>Week 1:</h4><br />
<p>Experimental:</p><br />
<ul><li>Expression of melittin</li><br />
<li>Checked protein expression by SDS-PAGE</li><br />
<li>Ligation of Rhl into mCherry plasmid</li><br />
<li> Double digestion to verify that ligation was successful</li></ul><br />
<p> Mathematical Modeling</p><br />
<ul><li>Adjusting model for protein expression</li><br />
<li>Visualization of melittin expression using MATLAB</li></ul><br />
<p>Outreach:</p><br />
<ul><li>Feature in Stony Brook Scholars Newsletter</li><br />
<li>Stony Brook involvement fair!</li></ul><br />
<br />
<h4>Week 2:</h4><br />
<p>Experimental:</p><ul><br />
<li>Measuring of fluroresence expression using TECAN </li><br />
<li>Found that Rhl is a leaky promoter, performed experiments to validate our concerns</li></ul><br />
<br />
<h4>Week 3</h4><br />
<ul><li>Culturing of cells with GST+TEV+Melittin</li><br />
<li>Checking protein expression by SDS-PAGE</li><br />
<li> Biobricking</li></ul><br />
</article><br />
</div><br />
</section> <br />
</div><br />
</body><br />
</html></div>
Hliu1423
http://2014.igem.org/Team:Stony_Brook/Notebook
Team:Stony Brook/Notebook
2014-10-17T23:32:57Z
<p>Hliu1423: </p>
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<div id="yellow_header"> <a href="http://www.stonybrook.edu/">Stony Brook University</a></div><br />
<div id="blue_header"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/1/1e/Stony_Brook_Apis-biotics_header.png"/ id="apis-biotics"/></a><br />
<div id="yellow_navigation_container"><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/7/78/Stony_Brook_NavIcon2_Home.png"/></a><br />
<p> Home </p><br />
</div><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook/Team"><img src="https://static.igem.org/mediawiki/2014/6/66/Stony_Brook_NavIcon2_Team.png"/></a><br />
<p> Team </p><br />
</div><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook/Project"><img src="https://static.igem.org/mediawiki/2014/9/99/Stony_Brook_NavIcon2_Project.png"/></a><br />
<p> Project </p><br />
</div><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook/Notebook"><img src="https://static.igem.org/mediawiki/2014/5/57/Stony_Brook_NavIcon2_Notebook.png"/></a><br />
<p> Notebook </p><br />
</div><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/8/80/Stony_Brook_NavIcon2_Outreach.png"/></a><br />
<p> Outreach </p><br />
</div><br />
<div id="yellow_navigation"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/0/03/Stony_Brook_NavIcon2_Acknowledgements.png"/></a><br />
<p> Attributions </p><br />
</div><br />
</div><br />
</div><br />
<div id="project"><br />
<p>Notebook</p><br />
</div><br />
<br />
<div class="picture"><img src="https://static.igem.org/mediawiki/2014/0/00/Stony_Brook_LabSpace.png" /></div><br />
<br />
</div><br />
<br />
<p class="glossary"><a href="https://2014.igem.org/Team:Stony_Brook/MaterialsMethods">Click here to see our procedures</a></p><br />
<br />
<div id="contents"> <section class="ac-container"><br />
<section class="ac-container"><br />
<div><br />
<input id="ac-1" name="accordion-1" type="checkbox" /><br />
<label for="ac-1">June</label><br />
<article class="ac-large"><br />
<br/><br/><h4>Week 1</h4><br />
<p>Literary research: </p><br />
<p>We looked into SUMO protein, a protective group that we could use to stabilize the melittin peptide. We also looked into mechanisms to express or release melittin:<br /><br />
<ul><li> Using a signalling sequence, signal peptides and pathways (OmpA, SecB, IgA beta)in order to secrete melittin</li><br />
<li>Lysing our cells with a killswitch and then using FLAG tags, affinity tags to purify melittin from lysate</li><br />
<li>Proteases to cleave off a protecting group.</li></ul></p> <br />
<p> We also looked into understanding biobricking assembly standards</p><br />
<br />
<h4>Week 2</h4> <br />
<p> Literary research:</p><br />
<ul> <li>Prepromelittin nucleotide sequence was found.</li><br />
<li> Decided on a fusion protein with melittin and GST tag</li><br />
<li>Reached out to Genomics Core Facility on Stony Brook University Campus to talk with Dr. John Schwedes about RNA extraction and Dr. Dmitri Gnatenko about primer design.</li><br />
<li>Searching for plasmids: met with Yelena from Molecular Cloning Facility to discuss appropriate plasmids for our experiment and search through their plasmid library</li></ul><br />
<p>Experimental:</p><br />
<ul><li>Competent E.coli cell line made. </li><br />
<li> Visited local beehive to obtain Apis mellifera samples; dissected our bees and extracted their RNA </li><br />
<li>Transformed RhlR biobrick</li></ul><br />
<p> Outreach:</p><br />
<p>We presented on synthetic biology at four classes at a local high school, East Sachem High School!</p><br />
<br />
<h4> Week 3</h4><br />
<p>Literary research:</p><br />
<ul><li> Looking into P. syringae as a model for P. aeruginosa?</li><br />
<li> Optimal expression conditions</li><br />
<li> Looking into TEV protease site to cleave off GST protein</li><br />
<li>Detection methods</li></ul><br />
<p>Experimental:</p><br />
<ul><li> Designed primers for melittin amplification and tagged melittin</li><br />
<li>Successful RNA extraction! (Picture from Week 7/2)</li><br />
<li>Obtained plasmid from our advisor, Dr. Czaplinski</li><br />
<li>Designed RhlR circuit and obtained signaling compound C4HSL</li><br />
<li>Constitutive promoter BBa_J23101 and J23102 transformed for later fluorescence testing</li></ul><br />
<br />
<h4>Week 4</h4><br />
<p>Experimental:</p><br />
<ul><li>Synthesized cDNA library</li><br />
<li> Designed and ordered Biobrick melittin primers</li><br />
<li>mCherry and fluorescence testing</li></ul><br />
<p>Outreach:</p><br />
<ul><li>Looked into doing an iGEM survey with other schools with the Stony Brook Institutional Review Board </li><br />
<li> Attended iGEM High School Jamboree! (picture) </li></ul><br />
</article><br />
</div><br />
<div><br />
<input id="ac-2" name="accordion-1" type="checkbox"><br />
<label for="ac-2">July</label><br />
<article class="ac-largeish"><br />
<br/><br/><h4>Week 1</h4><br />
<p>Experimental:</p><br />
<ul><li> Ligated and transformed for GST-melittin fusion protein</li><br />
<li>Induced plasmid with IPTG and checked with SDS page gel- no protein detected </li><br />
<li>Redid PCR and digest</li><br />
<li>Sequenced RhlR/Rhl circuit samples</li></ul><br />
<br />
<h4>Week 2</h4><br />
<p>Experimental:</p><br />
<ul><li>Prepared insert for our melittin plasmid- used PCR to amplify melittin, insert TEV cleavage site, then digested insert, and purified</li><br />
<li>Digested RhlR/ rhl plasmid, phosphatased, purified, ligated, and transformed. Cultured overnight, miniprepped and ran on gel to check results.</li><br />
<li>RhlR + Promoter miniprep, sent for sequencing</li></ul><br />
<p>Outreach</p><br />
<p>Presentated at Stony Brook University Biotechnology Camp for high school students!</p><br />
<h4>Week 3</h4><br />
<p>Experimental:</p><br />
<ul><li>Digest check, sequencing results for RhlR/Rhl</li><br />
<li>Screened colonies for melittin </li><br />
<li>Began PCR from beginning to amplify prepromelittin</li><br />
add-on PCR, inserted into duet plasmid</li></ul><br />
<p> Mathematical modeling:</p><br />
<p>Researching articles for rates of RhlR degradation</p><br />
<p>Comparison of our model with arbitrary values to experimental model for fluorescence</p><br />
<br />
<br />
<h4>Week 4</h4><br />
<p>Experimental:</p><br />
<ul><li>PCR amplification of prepromelittin from cDNA, addition of TEV site to melittin gene</li><br />
<li>Ligation of TEV-melittin into GST plasmid</li><br />
<li>Transformation of E.coli with ligation</li><br />
<li>Checked successful transformation by digesting transformed plasmid and viewing diegest on gel to cross-check with expected band size-70% success! </li><br />
<li>Samples from transformations sent for senquencing</li><br />
<li>N17+RhlR ligation into duet plasmid troubleshooting</li></ul><br />
<p> Mathematical modeling:</p><br />
<ul><li>Rates for C4HSL and RhlR degradation, dimerization</li><br />
<li>Preliminary mathematical models created!</li></ul><br />
<br />
</article><br />
</div><br />
<div><br />
<input id="ac-3" name="accordion-1" type="checkbox"><br />
<label for="ac-3">August</label><br />
<article class="ac-small"><br />
<br/><br/> <h4>Week 1</h4><br />
<p>Experimental:</p><br />
<ul><li>Results from sequencing: 3/4 samples were in frame; 1 had a missene mutation</li><br />
<li>Inducing of cultures of GST plasmid (control) and GST+TEV+melittin with IPTG</li><br />
<li> Attempted to checked protein expression by SDS-page</li><br />
<li>Co-transformations of RhlR plasmids</li></ul><br />
<h4>Week 2</h4><br />
<ul><li>Expression of melittin confirmed by SDS-PAGE!</li><br />
<li>Completion of Rhl circuit</li><br />
<li>Samples sent in for sequencing</li><br />
</ul><br />
<h4>Week 3</h4><br />
<p>School starts! </p><br />
</article><br />
</div><br />
<div><br />
<input id="ac-4" name="accordion-1" type="checkbox"><br />
<label for="ac-4">September</label><br />
<article class="ac-med"><br />
<br/><br/> <h4>Week 1:</h4><br />
<p>Experimental:</p><br />
<ul><li>Expression of melittin</li><br />
<li>Checked protein expression by SDS-PAGE</li><br />
<li>Ligation of Rhl into mCherry plasmid</li><br />
<li> Double digestion to verify that ligation was successful</li></ul><br />
<p> Mathematical Modeling</p><br />
<ul><li>Adjusting model for protein expression</li><br />
<li>Visualization of melittin expression using MATLAB</li></ul><br />
<p>Outreach:</p><br />
<ul><li>Feature in Stony Brook Scholars Newsletter</li><br />
<li>Stony Brook involvement fair!</li></ul><br />
<br />
<h4>Week 2:</h4><br />
<p>Experimental:</p><ul><br />
<li>Measuring of fluroresence expression using TECAN </li><br />
<li>Found that Rhl is a leaky promoter, performed experiments to validate our concerns</li></ul><br />
<br />
<h4>Week 3</h4><br />
<ul><li>Culturing of cells with GST+TEV+Melittin</li><br />
<li>Checking protein expression by SDS-PAGE</li><br />
<li> Biobricking</li></ul><br />
</article><br />
</div><br />
</section> <br />
</div><br />
</body><br />
</html></div>
Hliu1423
http://2014.igem.org/Team:Stony_Brook
Team:Stony Brook
2014-08-14T16:57:29Z
<p>Hliu1423: </p>
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<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Team"><img src="https://static.igem.org/mediawiki/2014/4/4a/Stony_Brook_NavIcon.Team.png" border="0px" /></a><br />
<p>Team</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Project"><img src="https://static.igem.org/mediawiki/2014/a/ab/Stony_Brook_NavIcon.Project.png" border="0px" /></a><br />
<p>Project</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Notebook"><img src="https://static.igem.org/mediawiki/2014/b/b5/Stony_Brook_NavIcon.Notebook.png" border="0px"/></a><br />
<p>Notebook</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="http://www.igem.org"> <img src="https://static.igem.org/mediawiki/2014/e/e2/Stony_Brook_Navicon.Outreach.png" border="0px"/></a><br />
<p>Outreach</p><br />
</div> <br />
<div id="navigation"><br />
<a href="http://www.igem.org"> <img src="https://static.igem.org/mediawiki/2014/5/5e/Stony_Brook_NavIcon.Acknowledgements.png" border="0px"/></a><br />
<p>Attributions</p><br />
</div> <br />
</div> --><br />
<br />
<div id="login"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook">page</a></li><br />
<li><a href="https://2014.igem.org/Talk:Team:Stony_Brook">discussion</a></li><br />
<li><a href="https://2014.igem.org/wiki/index.php?title=Team:Stony_Brook&amp;action=edit">view source</a></li><br />
<li><a href="https://2014.igem.org/wiki/index.php?title=Team:Stony_Brook&amp;action=history">history</a></li><br />
</ul><br />
</div><br />
<br />
<div id="background"><br />
</div><br />
<br />
<div id="projectDescription"><br />
<h1> Project Description </h1><br />
<p> The threat of antibiotic resistance has been a concern since the discovery of penicillin. As antibiotics continue to be used to treat disease, pathogenic bacteria continue to develop resistance, eventually becoming irresponsive to multiple classes of antibiotics. This is a major problem for areas that must maintain sterile environments, such as hospitals or other health-care settings. Hospital-acquired infections (HAI) are already an existing issue that poses risks to people receiving or recovering from treatment. Gram-negative bacteria in particular are known to cause more HAIs as they are more resistant to antibiotics than gram-positive bacteria. Irresponsible use of antibiotics has only exacerbated this issue. Recent reports from the World Health Organization confirm that antibiotic resistance is no longer something to worry about in the future, but a current, global health threat. </p><br />
<br />
<p> Alternatives to modern antibiotics, such as antimicrobial peptides, are now being researched. Antimicrobial peptides, or AMPs, are peptides produced innately by the immune systems of certain organisms to fight off infection. Because they kill microorganisms through different mechanisms, they have the potential to be effective against antibiotic resistant bacteria. One such AMP is melittin, produced by honey bees and used in their venom. Melittin kills cells by forming pores in the cell membrane, eventually destabilizing the cell and causing cell lysis. Because melittin targets a conserved area of cells, bacteria cannot develop resistance to it as easily as different types of antibiotics. </p><br />
<br />
<p> Our project this summer is to transform E. coli with the melittin gene in honey bees, so that the E. coli may express the melittin peptide, thereby developing a means to mass-produce melittin as an alternative to today's antibiotics. We hope to optimize the production of melittin in E. coli, as well as to test the bioactivity of our produced melittin in comparison to its naturally-occurring counterpart. </p><br />
</div><br />
</body><br />
<br />
</html></div>
Hliu1423
http://2014.igem.org/Team:Stony_Brook
Team:Stony Brook
2014-08-14T16:53:43Z
<p>Hliu1423: </p>
<hr />
<div><html xmlns="http://www.w3.org/1999/xhtml"><br />
<head><br />
<title>Stony Brook iGEM Homepage</title><br />
<meta http-equiv="Content-Type" content="text/html; charset=utf-8" /><br />
</head><br />
<body><br />
<a href="https://igem.org/Main_Page"><img src="https://static.igem.org/mediawiki/2014/5/51/Stony_brook_igem_logo.png" alt= "iGEM logo" name="iGEM_logo" border="0px" id="iGEM_logo" /></a><br />
<br />
<a href="https://www.facebook.com/iGEMatstonybrook"><img src="https://static.igem.org/mediawiki/2014/0/0f/Stony_brook_fb_logo.png" alt= "facebook" name="facebook" border="0px" id="facebook" /></a></a><br />
<a href="https://twitter.com/iGEMSBU"><img src="https://static.igem.org/mediawiki/2014/b/b5/Stony_brook_twitter_logo.png" alt= "twitter" name="twitter" border="0px" id="twitter" /></a></a><br />
<br />
<br />
<div id="header"><a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/1/11/Stony_brook_header.png" alt="Stony Brook iGEM" border="0px"" /></a></div><br />
<br />
<!-- <div id="navigation_container"><br />
<br />
<div id="navigation"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/b/b8/Stony_Brook_NavIcon.Home.png" border="0px" /></a><br />
<p>Home</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Team"><img src="https://static.igem.org/mediawiki/2014/4/4a/Stony_Brook_NavIcon.Team.png" border="0px" /></a><br />
<p>Team</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Project"><img src="https://static.igem.org/mediawiki/2014/a/ab/Stony_Brook_NavIcon.Project.png" border="0px" /></a><br />
<p>Project</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Notebook"><img src="https://static.igem.org/mediawiki/2014/b/b5/Stony_Brook_NavIcon.Notebook.png" border="0px"/></a><br />
<p>Notebook</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="http://www.igem.org"> <img src="https://static.igem.org/mediawiki/2014/e/e2/Stony_Brook_Navicon.Outreach.png" border="0px"/></a><br />
<p>Outreach</p><br />
</div> <br />
<div id="navigation"><br />
<a href="http://www.igem.org"> <img src="https://static.igem.org/mediawiki/2014/5/5e/Stony_Brook_NavIcon.Acknowledgements.png" border="0px"/></a><br />
<p>Attributions</p><br />
</div> <br />
</div> --><br />
<br />
<div id="login"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook">page</a></li><br />
<li><a href="https://2014.igem.org/Talk:Team:Stony_Brook">discussion</a></li><br />
<li><a href="https://2014.igem.org/wiki/index.php?title=Team:Stony_Brook&amp;action=edit">view source</a></li><br />
<li><a href="https://2014.igem.org/wiki/index.php?title=Team:Stony_Brook&amp;action=history">history</a></li><br />
</ul><br />
</div><br />
<br />
<div id="background"><br />
</div><br />
<br />
<div id="projectDescription"><br />
<h1> Project Description </h1><br />
<p> The threat of antibiotic resistance has been a concern since the discovery of penicillin. As antibiotics continue to be used to treat disease, pathogenic bacteria continue to develop resistance, eventually becoming irresponsive to multiple classes of antibiotics. This is a major problem for areas that must maintain sterile environments, such as hospitals or other health-care settings. Hospital-acquired infections (HAI) are already an existing issue that poses risks to people receiving or recovering from treatment. Gram-negative bacteria in particular are known to cause more HAIs as they are more resistant to antibiotics than gram-positive bacteria. Irresponsible use of antibiotics has only exacerbated this issue. Recent reports from the World Health Organization confirm that antibiotic resistance is no longer something to worry about in the future, but a current, global health threat. </p><br />
<br />
<p> Alternatives to modern antibiotics, such as antimicrobial peptides, are now being researched. Antimicrobial peptides, or AMPs, are peptides produced innately by the immune systems of certain organisms to fight off infection. Because they kill microorganisms through different mechanisms, they have the potential to be effective against antibiotic resistant bacteria. One such AMP is melittin, produced by honey bees and used in their venom. Melittin kills cells by forming pores in the cell membrane, eventually destabilizing the cell and causing cell lysis. Because melittin targets a conserved area of cells, bacteria cannot develop resistance to it as easily as different types of antibiotics. </p><br />
<br />
<p> Our project this summer is to transform E. coli with the melittin gene in honey bees, so that the E. coli may express the melittin peptide, thereby developing a means to mass-produce melittin as an alternative to today's antibiotics. We hope to optimize the production of melittin in E. coli, as well as to test the bioactivity of our produced melittin in comparison to its naturally-occurring counterpart. </p><br />
</div><br />
</body><br />
<br />
<style><br />
#contentSub, #search-controls, .firstHeading, #footer-box, #catlinks, #p-logo {<br />
display:none;}<br />
#top-section {<br />
border: none;<br />
height: 0px;}<br />
#content {<br />
border: none;}<br />
<br />
<br />
body {<br />
background-image: url(https://static.igem.org/mediawiki/2014/a/a5/Stony_brook_home_background.png);<br />
background-repeat:no-repeat;<br />
background-position: center center;<br />
background-attachment: fixed;<br />
-webkit-background-size: cover;<br />
-moz-background-size: cover;<br />
-o-background-size: cover;<br />
background-size: cover;<br />
}<br />
#header {<br />
width: 672px;<br />
position: absolute;<br />
top:29%;<br />
left:50%;<br />
margin-left: -310px;<br />
}<br />
<br />
#navigation {<br />
width: 65px;<br />
padding: 9px;<br />
float: left;<br />
color: #FFF;<br />
font-family:"Trebuchet MS", Arial, Helvetica, sans-serif;<br />
text-align: center;<br />
font-size:small;<br />
font-weight:bold;<br />
}<br />
<br />
#navigation p{<br />
display:none;<br />
}<br />
<br />
#navigation a:hover + p{<br />
display: block;<br />
}<br />
<br />
img:hover{<br />
opacity: 0.8;<br />
filter: alpha(opacity=80);<br />
}<br />
<br />
#iGEM_logo {<br />
position: absolute;<br />
left: 90%;<br />
top: 2%;<br />
}<br />
<br />
#navigation_container {<br />
width: 520 px;<br />
position: absolute;<br />
top: 40%;<br />
right: 50%;<br />
margin-right: -263px;<br />
}<br />
<br />
<br />
a{<br />
outline: 0;<br />
}<br />
<br />
img{<br />
border: 0;<br />
}<br />
<br />
#login{<br />
width: 400px;<br />
height: 20px;<br />
background-color: #000;<br />
opacity: 0.7;<br />
filter: alpha(opacity=70);<br />
position: absolute;<br />
top: 0;<br />
left:0;<br />
}<br />
#login ul{<br />
list-style-type: none;<br />
margin: 0;<br />
padding: 0px;<br />
}<br />
<br />
#login li{<br />
float: left;<br />
text-align: center;<br />
}<br />
<br />
#login a{<br />
display: block;<br />
height: 20px;<br />
line-height: 20px;<br />
width: 100px;<br />
text-decoration: none;<br />
color: #FFF;<br />
font-family: "Trebuchet MS", Arial, Helvetica, sans-serif;<br />
font-size: x-small;<br />
vertical-align: middle;<br />
font-weight: bold;<br />
color: #FFF;<br />
}<br />
<br />
#login a:hover{<br />
color: #0F0;<br />
}<br />
<br />
#facebook{<br />
position: absolute;<br />
bottom: 1%;<br />
left: 1%;<br />
padding-bottom: 5px;<br />
}<br />
<br />
#twitter{<br />
position: absolute;<br />
bottom: 1%;<br />
left: 7%;<br />
padding-bottom: 5px;<br />
}<br />
<br />
</style><br />
</html></div>
Hliu1423
http://2014.igem.org/Team:Stony_Brook
Team:Stony Brook
2014-08-14T16:51:39Z
<p>Hliu1423: </p>
<hr />
<div><html xmlns="http://www.w3.org/1999/xhtml"><br />
<head><br />
<title>Stony Brook iGEM Homepage</title><br />
<meta http-equiv="Content-Type" content="text/html; charset=utf-8" /><br />
</head><br />
<body><br />
<a href="https://igem.org/Main_Page"><img src="https://static.igem.org/mediawiki/2014/5/51/Stony_brook_igem_logo.png" alt= "iGEM logo" name="iGEM_logo" border="0px" id="iGEM_logo" /></a><br />
<br />
<a href="https://www.facebook.com/iGEMatstonybrook"><img src="https://static.igem.org/mediawiki/2014/0/0f/Stony_brook_fb_logo.png" alt= "facebook" name="facebook" border="0px" id="facebook" /></a></a><br />
<a href="https://twitter.com/iGEMSBU"><img src="https://static.igem.org/mediawiki/2014/b/b5/Stony_brook_twitter_logo.png" alt= "twitter" name="twitter" border="0px" id="twitter" /></a></a><br />
<br />
<br />
<div id="header"><a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/1/11/Stony_brook_header.png" alt="Stony Brook iGEM" border="0px"" /></a></div><br />
<br />
<!-- <div id="navigation_container"><br />
<br />
<div id="navigation"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/b/b8/Stony_Brook_NavIcon.Home.png" border="0px" /></a><br />
<p>Home</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Team"><img src="https://static.igem.org/mediawiki/2014/4/4a/Stony_Brook_NavIcon.Team.png" border="0px" /></a><br />
<p>Team</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Project"><img src="https://static.igem.org/mediawiki/2014/a/ab/Stony_Brook_NavIcon.Project.png" border="0px" /></a><br />
<p>Project</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Notebook"><img src="https://static.igem.org/mediawiki/2014/b/b5/Stony_Brook_NavIcon.Notebook.png" border="0px"/></a><br />
<p>Notebook</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="http://www.igem.org"> <img src="https://static.igem.org/mediawiki/2014/e/e2/Stony_Brook_Navicon.Outreach.png" border="0px"/></a><br />
<p>Outreach</p><br />
</div> <br />
<div id="navigation"><br />
<a href="http://www.igem.org"> <img src="https://static.igem.org/mediawiki/2014/5/5e/Stony_Brook_NavIcon.Acknowledgements.png" border="0px"/></a><br />
<p>Attributions</p><br />
</div> <br />
</div> --><br />
<br />
<div id="login"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook">page</a></li><br />
<li><a href="https://2014.igem.org/Talk:Team:Stony_Brook">discussion</a></li><br />
<li><a href="https://2014.igem.org/wiki/index.php?title=Team:Stony_Brook&amp;action=edit">view source</a></li><br />
<li><a href="https://2014.igem.org/wiki/index.php?title=Team:Stony_Brook&amp;action=history">history</a></li><br />
</ul><br />
</div><br />
<br />
<div id="background"><br />
</div><br />
<br />
<div id="projectDescription"><br />
<h1> Project Description </h1><br />
<p> The threat of antibiotic resistance has been a concern since the discovery of penicillin. As antibiotics continue to be used to treat disease, pathogenic bacteria continue to develop resistance, eventually becoming irresponsive to multiple classes of antibiotics. This is a major problem for areas that must maintain sterile environments, such as hospitals or other health-care settings. Hospital-acquired infections (HAI) are already an existing issue that poses risks to people receiving or recovering from treatment. Gram-negative bacteria in particular are known to cause more HAIs as they are more resistant to antibiotics than gram-positive bacteria. Irresponsible use of antibiotics has only exacerbated this issue. Recent reports from the World Health Organization confirm that antibiotic resistance is no longer something to worry about in the future, but a current, global health threat. </p><br />
<br />
<p> Alternatives to modern antibiotics, such as antimicrobial peptides, are now being researched. Antimicrobial peptides, or AMPs, are peptides produced innately by the immune systems of certain organisms to fight off infection. Because they kill microorganisms through different mechanisms, they have the potential to be effective against antibiotic resistant bacteria. One such AMP is melittin, produced by honey bees and used in their venom. Melittin kills cells by forming pores in the cell membrane, eventually destabilizing the cell and causing cell lysis. Because melittin targets a conserved area of cells, bacteria cannot develop resistance to it as easily as different types of antibiotics. </p><br />
<br />
<p> Our project this summer is to transform E. coli with the melittin gene in honey bees, so that the E. coli may express the melittin peptide, thereby developing a means to mass-produce melittin as an alternative to today's antibiotics. We hope to optimize the production of melittin in E. coli, as well as to test the bioactivity of our produced melittin in comparison to its naturally-occurring counterpart. </p><br />
</div><br />
<br />
<br />
<style><br />
#contentSub, #search-controls, .firstHeading, #footer-box, #catlinks, #p-logo {<br />
display:none;}<br />
#top-section {<br />
border: none;<br />
height: 0px;}<br />
#content {<br />
border: none;}<br />
</body><br />
<br />
body {<br />
background-image: url(home%20page%20background.png);<br />
background-repeat:no-repeat;<br />
background-position: center center;<br />
background-attachment: fixed;<br />
-webkit-background-size: cover;<br />
-moz-background-size: cover;<br />
-o-background-size: cover;<br />
background-size: cover;<br />
}<br />
#header {<br />
width: 672px;<br />
position: absolute;<br />
top:29%;<br />
left:50%;<br />
margin-left: -310px;<br />
}<br />
<br />
#navigation {<br />
width: 65px;<br />
padding: 9px;<br />
float: left;<br />
color: #FFF;<br />
font-family:"Trebuchet MS", Arial, Helvetica, sans-serif;<br />
text-align: center;<br />
font-size:small;<br />
font-weight:bold;<br />
}<br />
<br />
#navigation p{<br />
display:none;<br />
}<br />
<br />
#navigation a:hover + p{<br />
display: block;<br />
}<br />
<br />
img:hover{<br />
opacity: 0.8;<br />
filter: alpha(opacity=80);<br />
}<br />
<br />
#iGEM_logo {<br />
position: absolute;<br />
left: 90%;<br />
top: 2%;<br />
}<br />
<br />
#navigation_container {<br />
width: 520 px;<br />
position: absolute;<br />
top: 40%;<br />
right: 50%;<br />
margin-right: -263px;<br />
}<br />
<br />
<br />
a{<br />
outline: 0;<br />
}<br />
<br />
img{<br />
border: 0;<br />
}<br />
<br />
#login{<br />
width: 400px;<br />
height: 20px;<br />
background-color: #000;<br />
opacity: 0.7;<br />
filter: alpha(opacity=70);<br />
position: absolute;<br />
top: 0;<br />
left:0;<br />
}<br />
#login ul{<br />
list-style-type: none;<br />
margin: 0;<br />
padding: 0px;<br />
}<br />
<br />
#login li{<br />
float: left;<br />
text-align: center;<br />
}<br />
<br />
#login a{<br />
display: block;<br />
height: 20px;<br />
line-height: 20px;<br />
width: 100px;<br />
text-decoration: none;<br />
color: #FFF;<br />
font-family: "Trebuchet MS", Arial, Helvetica, sans-serif;<br />
font-size: x-small;<br />
vertical-align: middle;<br />
font-weight: bold;<br />
color: #FFF;<br />
}<br />
<br />
#login a:hover{<br />
color: #0F0;<br />
}<br />
<br />
#facebook{<br />
position: absolute;<br />
bottom: 1%;<br />
left: 1%;<br />
padding-bottom: 5px;<br />
}<br />
<br />
#twitter{<br />
position: absolute;<br />
bottom: 1%;<br />
left: 7%;<br />
padding-bottom: 5px;<br />
}<br />
<br />
</style><br />
</html></div>
Hliu1423
http://2014.igem.org/Team:Stony_Brook
Team:Stony Brook
2014-08-14T16:46:28Z
<p>Hliu1423: </p>
<hr />
<div><html xmlns="http://www.w3.org/1999/xhtml"><br />
<head><br />
<title>Stony Brook iGEM Homepage</title><br />
<meta http-equiv="Content-Type" content="text/html; charset=utf-8" /><br />
</head><br />
<body><br />
</body><br />
<style><br />
#contentSub, #search-controls, .firstHeading, #footer-box, #catlinks, #p-logo {<br />
display:none;}<br />
#top-section {<br />
border: none;<br />
height: 0px;}<br />
#content {<br />
border: none;}<br />
<br />
</style></div>
Hliu1423
http://2014.igem.org/Team:Stony_Brook
Team:Stony Brook
2014-08-14T16:29:54Z
<p>Hliu1423: </p>
<hr />
<div>{{CSS/Main}}</div>
Hliu1423
http://2014.igem.org/Talk:Team:Stony_Brook/Project
Talk:Team:Stony Brook/Project
2014-08-14T16:29:00Z
<p>Hliu1423: Created page with "{{CSS/Main}}"</p>
<hr />
<div>{{CSS/Main}}</div>
Hliu1423
http://2014.igem.org/Team:Stony_Brook
Team:Stony Brook
2014-08-14T16:23:49Z
<p>Hliu1423: Blanked the page</p>
<hr />
<div></div>
Hliu1423
http://2014.igem.org/Team:Stony_Brook
Team:Stony Brook
2014-08-14T16:20:31Z
<p>Hliu1423: </p>
<hr />
<div><head><br />
<title>Stony Brook iGEM Homepage</title><br />
<meta http-equiv="Content-Type" content="text/html; charset=utf-8" /><br />
<br />
<html><br />
</body><br />
<style type="text/css"><br />
body {<br />
background-image: url(https://static.igem.org/mediawiki/2014/a/a5/Stony_brook_home_background.png);<br />
background-repeat:no-repeat;<br />
background-position: center center;<br />
background-attachment: fixed;<br />
-webkit-background-size: cover;<br />
-moz-background-size: cover;<br />
-o-background-size: cover;<br />
background-size: cover;<br />
}<br />
#header {<br />
width: 672px;<br />
position: absolute;<br />
top:29%;<br />
left:50%;<br />
margin-left: -310px;<br />
margin-top:-50px;<br />
}<br />
<br />
#navigation {<br />
width: 65px;<br />
padding: 9px;<br />
float: left;<br />
color: #FFF;<br />
font-family:"Trebuchet MS", Arial, Helvetica, sans-serif;<br />
text-align: center;<br />
font-size:small;<br />
font-weight:bold;<br />
}<br />
<br />
#navigation p{<br />
display:none;<br />
}<br />
<br />
#navigation a:hover + p{<br />
display: block;<br />
}<br />
<br />
img:hover{<br />
opacity: 0.8;<br />
filter: alpha(opacity=80);<br />
}<br />
<br />
#iGEM_logo {<br />
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<br />
<a href="https://igem.org/Main_Page"><img src="https://static.igem.org/mediawiki/2014/5/51/Stony_brook_igem_logo.png" alt= "iGEM logo" name="iGEM_logo" border="0px" id="iGEM_logo" /></a><br />
<br />
<a href="https://www.facebook.com/iGEMatstonybrook"><img src="https://static.igem.org/mediawiki/2014/0/0f/Stony_brook_fb_logo.png" alt= "facebook" name="facebook" border="0px" id="facebook" /></a></a><br />
<a href="https://twitter.com/iGEMSBU"><img src="https://static.igem.org/mediawiki/2014/b/b5/Stony_brook_twitter_logo.png" alt= "twitter" name="twitter" border="0px" id="twitter" /></a></a><br />
<br />
<br />
<div id="header"><a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/1/11/Stony_brook_header.png" alt="Stony Brook iGEM" border="0px"" /></a></div><br />
<br />
<!-- <div id="navigation_container"><br />
<br />
<div id="navigation"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/b/b8/Stony_Brook_NavIcon.Home.png" border="0px" /></a><br />
<p>Home</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Team"><img src="https://static.igem.org/mediawiki/2014/4/4a/Stony_Brook_NavIcon.Team.png" border="0px" /></a><br />
<p>Team</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Project"><img src="https://static.igem.org/mediawiki/2014/a/ab/Stony_Brook_NavIcon.Project.png" border="0px" /></a><br />
<p>Project</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Notebook"><img src="https://static.igem.org/mediawiki/2014/b/b5/Stony_Brook_NavIcon.Notebook.png" border="0px"/></a><br />
<p>Notebook</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="http://www.igem.org"> <img src="https://static.igem.org/mediawiki/2014/e/e2/Stony_Brook_Navicon.Outreach.png" border="0px"/></a><br />
<p>Outreach</p><br />
</div> <br />
<div id="navigation"><br />
<a href="http://www.igem.org"> <img src="https://static.igem.org/mediawiki/2014/5/5e/Stony_Brook_NavIcon.Acknowledgements.png" border="0px"/></a><br />
<p>Attributions</p><br />
</div> <br />
</div> --><br />
<br />
<div id="login"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook">page</a></li><br />
<li><a href="https://2014.igem.org/Talk:Team:Stony_Brook">discussion</a></li><br />
<li><a href="https://2014.igem.org/wiki/index.php?title=Team:Stony_Brook&amp;action=edit">view source</a></li><br />
<li><a href="https://2014.igem.org/wiki/index.php?title=Team:Stony_Brook&amp;action=history">history</a></li><br />
</ul><br />
</div><br />
<br />
<div id="background"><br />
</div><br />
<br />
<div id="projectDescription"><br />
<h1> Project Description </h1><br />
<p> The threat of antibiotic resistance has been a concern since the discovery of penicillin. As antibiotics continue to be used to treat disease, pathogenic bacteria continue to develop resistance, eventually becoming irresponsive to multiple classes of antibiotics. This is a major problem for areas that must maintain sterile environments, such as hospitals or other health-care settings. Hospital-acquired infections (HAI) are already an existing issue that poses risks to people receiving or recovering from treatment. Gram-negative bacteria in particular are known to cause more HAIs as they are more resistant to antibiotics than gram-positive bacteria. Irresponsible use of antibiotics has only exacerbated this issue. Recent reports from the World Health Organization confirm that antibiotic resistance is no longer something to worry about in the future, but a current, global health threat. </p><br />
<br />
<p> Alternatives to modern antibiotics, such as antimicrobial peptides, are now being researched. Antimicrobial peptides, or AMPs, are peptides produced innately by the immune systems of certain organisms to fight off infection. Because they kill microorganisms through different mechanisms, they have the potential to be effective against antibiotic resistant bacteria. One such AMP is melittin, produced by honey bees and used in their venom. Melittin kills cells by forming pores in the cell membrane, eventually destabilizing the cell and causing cell lysis. Because melittin targets a conserved area of cells, bacteria cannot develop resistance to it as easily as different types of antibiotics. </p><br />
<br />
<p> Our project this summer is to transform E. coli with the melittin gene in honey bees, so that the E. coli may express the melittin peptide, thereby developing a means to mass-produce melittin as an alternative to today's antibiotics. We hope to optimize the production of melittin in E. coli, as well as to test the bioactivity of our produced melittin in comparison to its naturally-occurring counterpart. </p><br />
</div><br />
<br />
</html></div>
Hliu1423
http://2014.igem.org/Team:Stony_Brook
Team:Stony Brook
2014-08-14T16:15:08Z
<p>Hliu1423: </p>
<hr />
<div><head><br />
<title>Stony Brook iGEM Homepage</title><br />
<meta http-equiv="Content-Type" content="text/html; charset=utf-8" /><br />
<br />
<br />
</body><br />
<style type="text/css"><br />
body {<br />
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<br />
<br />
<br />
<a href="https://igem.org/Main_Page"><img src="https://static.igem.org/mediawiki/2014/5/51/Stony_brook_igem_logo.png" alt= "iGEM logo" name="iGEM_logo" border="0px" id="iGEM_logo" /></a><br />
<br />
<a href="https://www.facebook.com/iGEMatstonybrook"><img src="https://static.igem.org/mediawiki/2014/0/0f/Stony_brook_fb_logo.png" alt= "facebook" name="facebook" border="0px" id="facebook" /></a></a><br />
<a href="https://twitter.com/iGEMSBU"><img src="https://static.igem.org/mediawiki/2014/b/b5/Stony_brook_twitter_logo.png" alt= "twitter" name="twitter" border="0px" id="twitter" /></a></a><br />
<br />
<br />
<div id="header"><a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/1/11/Stony_brook_header.png" alt="Stony Brook iGEM" border="0px"" /></a></div><br />
<br />
<!-- <div id="navigation_container"><br />
<br />
<div id="navigation"> <a href="https://2014.igem.org/Team:Stony_Brook"><img src="https://static.igem.org/mediawiki/2014/b/b8/Stony_Brook_NavIcon.Home.png" border="0px" /></a><br />
<p>Home</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Team"><img src="https://static.igem.org/mediawiki/2014/4/4a/Stony_Brook_NavIcon.Team.png" border="0px" /></a><br />
<p>Team</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Project"><img src="https://static.igem.org/mediawiki/2014/a/ab/Stony_Brook_NavIcon.Project.png" border="0px" /></a><br />
<p>Project</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="https://2014.igem.org/Team:Stony_Brook/Notebook"><img src="https://static.igem.org/mediawiki/2014/b/b5/Stony_Brook_NavIcon.Notebook.png" border="0px"/></a><br />
<p>Notebook</p><br />
</div><br />
<br />
<div id="navigation"><br />
<a href="http://www.igem.org"> <img src="https://static.igem.org/mediawiki/2014/e/e2/Stony_Brook_Navicon.Outreach.png" border="0px"/></a><br />
<p>Outreach</p><br />
</div> <br />
<div id="navigation"><br />
<a href="http://www.igem.org"> <img src="https://static.igem.org/mediawiki/2014/5/5e/Stony_Brook_NavIcon.Acknowledgements.png" border="0px"/></a><br />
<p>Attributions</p><br />
</div> <br />
</div> --><br />
<br />
<div id="login"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook">page</a></li><br />
<li><a href="https://2014.igem.org/Talk:Team:Stony_Brook">discussion</a></li><br />
<li><a href="https://2014.igem.org/wiki/index.php?title=Team:Stony_Brook&amp;action=edit">view source</a></li><br />
<li><a href="https://2014.igem.org/wiki/index.php?title=Team:Stony_Brook&amp;action=history">history</a></li><br />
</ul><br />
</div><br />
<br />
<div id="background"><br />
</div><br />
<br />
<div id="projectDescription"><br />
<h1> Project Description </h1><br />
<p> The threat of antibiotic resistance has been a concern since the discovery of penicillin. As antibiotics continue to be used to treat disease, pathogenic bacteria continue to develop resistance, eventually becoming irresponsive to multiple classes of antibiotics. This is a major problem for areas that must maintain sterile environments, such as hospitals or other health-care settings. Hospital-acquired infections (HAI) are already an existing issue that poses risks to people receiving or recovering from treatment. Gram-negative bacteria in particular are known to cause more HAIs as they are more resistant to antibiotics than gram-positive bacteria. Irresponsible use of antibiotics has only exacerbated this issue. Recent reports from the World Health Organization confirm that antibiotic resistance is no longer something to worry about in the future, but a current, global health threat. </p><br />
<br />
<p> Alternatives to modern antibiotics, such as antimicrobial peptides, are now being researched. Antimicrobial peptides, or AMPs, are peptides produced innately by the immune systems of certain organisms to fight off infection. Because they kill microorganisms through different mechanisms, they have the potential to be effective against antibiotic resistant bacteria. One such AMP is melittin, produced by honey bees and used in their venom. Melittin kills cells by forming pores in the cell membrane, eventually destabilizing the cell and causing cell lysis. Because melittin targets a conserved area of cells, bacteria cannot develop resistance to it as easily as different types of antibiotics. </p><br />
<br />
<p> Our project this summer is to transform E. coli with the melittin gene in honey bees, so that the E. coli may express the melittin peptide, thereby developing a means to mass-produce melittin as an alternative to today's antibiotics. We hope to optimize the production of melittin in E. coli, as well as to test the bioactivity of our produced melittin in comparison to its naturally-occurring counterpart. </p><br />
</div><br />
<br />
</html></div>
Hliu1423
http://2014.igem.org/File:Team_Stonybrook_UG-Bio-banner.jpg
File:Team Stonybrook UG-Bio-banner.jpg
2014-06-30T15:50:18Z
<p>Hliu1423: </p>
<hr />
<div></div>
Hliu1423
http://2014.igem.org/Team:Stony_Brook
Team:Stony Brook
2014-06-30T15:42:00Z
<p>Hliu1423: </p>
<hr />
<div>{{CSS/Main}}<br />
{{DISPLAYTITLE:<span style="display: none"></span>}}<br />
<br />
<html><br />
<style><br />
#contentSub, #footer-box, #catlinks, #search-controls, #p-logo, .printfooter, .firstHeading,.visualClear {display: none;} /*-- hides default wiki settings --*/<br />
</style><br />
<!--main content --><br />
<table width="70%" align="center"><br />
<br />
<br />
<!--welcome box --><br />
<tr><br />
<td style="border:1px solid black;" colspan="3" align="center" height="150px" bgColor=#FF404B><br />
<h1 >WELCOME TO iGEM 2014! </h1><br />
<p>Your team has been approved and you are ready to start the iGEM season!<br />
<br>On this page you can document your project, introduce your team members, document your progress <br> and share your iGEM experience with the rest of the world! </p><br />
<br><br />
<p style="color:#E7E7E7"> <a href="https://2014.igem.org/wiki/index.php?title=Team:Stony_Brook&action=edit"style="color:#FFFFFF"> Click here to edit this page!</a> </p><br />
</td><br />
</tr><br />
<br />
<tr> <td colspan="3" height="5px"> </td></tr><br />
<!-- end welcome box --><br />
<br />
<tr> <br />
<br />
<!--navigation menu --><br />
<td align="center" colspan="3"><br />
<br />
<table width="100%"><br />
<tr heigth="15px"></tr><br />
<tr heigth="75px"> <br />
<br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook"style="color:#000000">Home </a> </td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Team"style="color:#000000"> Team </a> </td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://igem.org/Team.cgi?year=2014&team_name=Stony_Brook"style="color:#000000"> Official Team Profile </a></td><br />
<br />
<td style="border:1px solid black" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Project"style="color:#000000"> Project</a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Parts"style="color:#000000"> Parts</a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Modeling"style="color:#000000"> Modeling</a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Notebook"style="color:#000000"> Notebook</a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Safety"style=" color:#000000"> Safety </a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Attributions"style="color:#000000"> Attributions </a></td><br />
<br />
<br />
<td align ="center"> <a href="https://2014.igem.org/Main_Page"> <img src="https://static.igem.org/mediawiki/igem.org/6/60/Igemlogo_300px.png" width="55px"></a> </td><br />
</tr><br />
</table><br />
<br />
<!--end navigation menu --><br />
</tr><br />
<br />
<br />
</tr><br />
<br />
<br />
<br />
<br />
<br />
</td><br />
<br />
<tr> <td colspan="3" height="15px"> </td></tr><br />
<tr><td bgColor="#e7e7e7" colspan="3" height="1px"> </tr><br />
<tr> <td colspan="3" height="5px"> </td></tr><br />
<br />
<br />
<!--requirements section --><br />
<tr><td colspan="3"> <h3> Requirements </h3></td></tr><br />
<tr><br />
<td width="45%" valign="top"> <br />
<br />
<p> Please be sure to keep these links, your audience will want to find your: </p><br />
<br />
<!-- Links to other team pages --><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook">Home</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Team">Team</a> </li><br />
<li><a href="https://igem.org/Team.cgi?year=2013&team_name=Stony_Brook">Official Team Profile</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Project">Project</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Parts">Parts</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Modeling">Modeling</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Notebook">Notebook</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Safety">Safety</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Attributions">Attributions</a> </li><br />
<br />
</ul><br />
<br />
</td><br />
<br />
<td > </td><br />
<td width="45%"><br />
<br />
<p>There are a few wiki requirements teams must follow:</p><br />
<ul><br />
<li>All pages, images and files must be hosted on the <a href ="https://2014.igem.org/Special:Upload"> 2014.igem.org server</a>. </li><br />
<li>All pages must be created under the team’s name space.</li><br />
<li>As part of your documentation, keep the links from the menu to the left. </li><br />
<li>Do not use flash in wiki code. </li><br />
<li>The <a href="https://static.igem.org/mediawiki/igem.org/6/60/Igemlogo_300px.png"> iGEM logo </a> should be placed on the upper part of every page and should link to <a href="https://2014.igem.org/Main_Page">2014.igem.org</a>.</li><br />
</ul><br />
<p>Visit the <a href="https://2014.igem.org/Wiki_How-To"> Wiki How To page </a> for a complete list of requirements, tips and other useful information. </p><br />
<br />
</td><br />
</tr><br />
<br />
<br />
<tr> <td colspan="3" height="15px"> </td></tr><br />
<tr><td bgColor="#e7e7e7" colspan="3" height="1"> </tr><br />
<br />
<br />
<!--tips --><br />
<tr><td colspan="3" > <h3> Tips </h3></td></tr><br />
<br />
<tr><br />
<td width="45%" valign="top"> <br />
<p>We are currently working on providing teams with some easy to use design templates.<br />
<br> In the meantime you can also view other team wikis for inspiration! Here are some very good examples</p><br />
<br />
<ul><br />
<li> <a href="https://2013.igem.org/Team:SDU-Denmark/"> 2013 SDU Denmark </a> </li><br />
<li> <a href="https://2013.igem.org/Team:SYSU-China">2013 SYSU China</a> </li><br />
<li> <a href="https://2013.igem.org/Team:Shenzhen_BGIC_ATCG"> 2013 Shenxhen BGIG ATCG </a></li><br />
<li> <a href="https://2013.igem.org/Team:Colombia_Uniandes">2013 Colombia Unianades </a></li><br />
<li> <a href="https://2013.igem.org/Team:Lethbridge">2013 Lethbridge</a></li><br />
</ul><br />
<br />
<p>For a full wiki list, you can visit <a href="https://igem.org/Team_Wikis?year=2013">iGEM 2013 web sites </a> and <a href="https://igem.org/Team_Wikis?year=2012">iGEM 2012 web sites</a> lists. </p><br />
</td><br />
<br />
<td> </td><br />
<td width="45%"> <br />
<br />
<p>This wiki will be your team’s first interaction with the rest of the world, so here are a few tips to help you get started: </p><br />
<br />
<ul><br />
<li>State your accomplishments! Tell people what you have achieved from the start. </li><br />
<li>Be clear about what you are doing and what you plan to do.</li><br />
<li>You have a global audience! Consider the different backgrounds that your users come from.</li><br />
<li>Make sure information is easy to find; nothing should be more than 3 clicks away. </li><br />
<li>Avoid using very small fonts and low contrast colors; information should be easy to read. </li><br />
<li>Start documenting your project as early as possible; don’t leave anything to the last minute before the Wiki Freeze. For a complete list of deadlines visit the <a href="">iGEM 2013 calendar</a> </li><br />
<li>Have lots of fun! </li><br />
</ul><br />
</br><br />
</td><br />
</tr><br />
</table></div>
Hliu1423
http://2014.igem.org/Team:Stony_Brook
Team:Stony Brook
2014-06-30T15:36:05Z
<p>Hliu1423: </p>
<hr />
<div>{{CSS/Main}}<br />
{{DISPLAYTITLE:<span style="display: none"></span>}}<br />
<br />
<html><br />
<br />
<!--main content --><br />
<table width="70%" align="center"><br />
<br />
<br />
<!--welcome box --><br />
<tr><br />
<td style="border:1px solid black;" colspan="3" align="center" height="150px" bgColor=#FF404B><br />
<h1 >WELCOME TO iGEM 2014! </h1><br />
<p>Your team has been approved and you are ready to start the iGEM season!<br />
<br>On this page you can document your project, introduce your team members, document your progress <br> and share your iGEM experience with the rest of the world! </p><br />
<br><br />
<p style="color:#E7E7E7"> <a href="https://2014.igem.org/wiki/index.php?title=Team:Stony_Brook&action=edit"style="color:#FFFFFF"> Click here to edit this page!</a> </p><br />
</td><br />
</tr><br />
<br />
<tr> <td colspan="3" height="5px"> </td></tr><br />
<!-- end welcome box --><br />
<br />
<tr> <br />
<br />
<!--navigation menu --><br />
<td align="center" colspan="3"><br />
<br />
<table width="100%"><br />
<tr heigth="15px"></tr><br />
<tr heigth="75px"> <br />
<br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook"style="color:#000000">Home </a> </td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Team"style="color:#000000"> Team </a> </td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://igem.org/Team.cgi?year=2014&team_name=Stony_Brook"style="color:#000000"> Official Team Profile </a></td><br />
<br />
<td style="border:1px solid black" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Project"style="color:#000000"> Project</a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Parts"style="color:#000000"> Parts</a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Modeling"style="color:#000000"> Modeling</a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Notebook"style="color:#000000"> Notebook</a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Safety"style=" color:#000000"> Safety </a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Attributions"style="color:#000000"> Attributions </a></td><br />
<br />
<br />
<td align ="center"> <a href="https://2014.igem.org/Main_Page"> <img src="https://static.igem.org/mediawiki/igem.org/6/60/Igemlogo_300px.png" width="55px"></a> </td><br />
</tr><br />
</table><br />
<br />
<!--end navigation menu --><br />
</tr><br />
<br />
<br />
</tr><br />
<br />
<br />
<br />
<br />
<br />
</td><br />
<br />
<tr> <td colspan="3" height="15px"> </td></tr><br />
<tr><td bgColor="#e7e7e7" colspan="3" height="1px"> </tr><br />
<tr> <td colspan="3" height="5px"> </td></tr><br />
<br />
<br />
<!--requirements section --><br />
<tr><td colspan="3"> <h3> Requirements </h3></td></tr><br />
<tr><br />
<td width="45%" valign="top"> <br />
<br />
<p> Please be sure to keep these links, your audience will want to find your: </p><br />
<br />
<!-- Links to other team pages --><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook">Home</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Team">Team</a> </li><br />
<li><a href="https://igem.org/Team.cgi?year=2013&team_name=Stony_Brook">Official Team Profile</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Project">Project</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Parts">Parts</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Modeling">Modeling</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Notebook">Notebook</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Safety">Safety</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Attributions">Attributions</a> </li><br />
<br />
</ul><br />
<br />
</td><br />
<br />
<td > </td><br />
<td width="45%"><br />
<br />
<p>There are a few wiki requirements teams must follow:</p><br />
<ul><br />
<li>All pages, images and files must be hosted on the <a href ="https://2014.igem.org/Special:Upload"> 2014.igem.org server</a>. </li><br />
<li>All pages must be created under the team’s name space.</li><br />
<li>As part of your documentation, keep the links from the menu to the left. </li><br />
<li>Do not use flash in wiki code. </li><br />
<li>The <a href="https://static.igem.org/mediawiki/igem.org/6/60/Igemlogo_300px.png"> iGEM logo </a> should be placed on the upper part of every page and should link to <a href="https://2014.igem.org/Main_Page">2014.igem.org</a>.</li><br />
</ul><br />
<p>Visit the <a href="https://2014.igem.org/Wiki_How-To"> Wiki How To page </a> for a complete list of requirements, tips and other useful information. </p><br />
<br />
</td><br />
</tr><br />
<br />
<br />
<tr> <td colspan="3" height="15px"> </td></tr><br />
<tr><td bgColor="#e7e7e7" colspan="3" height="1"> </tr><br />
<br />
<br />
<!--tips --><br />
<tr><td colspan="3" > <h3> Tips </h3></td></tr><br />
<br />
<tr><br />
<td width="45%" valign="top"> <br />
<p>We are currently working on providing teams with some easy to use design templates.<br />
<br> In the meantime you can also view other team wikis for inspiration! Here are some very good examples</p><br />
<br />
<ul><br />
<li> <a href="https://2013.igem.org/Team:SDU-Denmark/"> 2013 SDU Denmark </a> </li><br />
<li> <a href="https://2013.igem.org/Team:SYSU-China">2013 SYSU China</a> </li><br />
<li> <a href="https://2013.igem.org/Team:Shenzhen_BGIC_ATCG"> 2013 Shenxhen BGIG ATCG </a></li><br />
<li> <a href="https://2013.igem.org/Team:Colombia_Uniandes">2013 Colombia Unianades </a></li><br />
<li> <a href="https://2013.igem.org/Team:Lethbridge">2013 Lethbridge</a></li><br />
</ul><br />
<br />
<p>For a full wiki list, you can visit <a href="https://igem.org/Team_Wikis?year=2013">iGEM 2013 web sites </a> and <a href="https://igem.org/Team_Wikis?year=2012">iGEM 2012 web sites</a> lists. </p><br />
</td><br />
<br />
<td> </td><br />
<td width="45%"> <br />
<br />
<p>This wiki will be your team’s first interaction with the rest of the world, so here are a few tips to help you get started: </p><br />
<br />
<ul><br />
<li>State your accomplishments! Tell people what you have achieved from the start. </li><br />
<li>Be clear about what you are doing and what you plan to do.</li><br />
<li>You have a global audience! Consider the different backgrounds that your users come from.</li><br />
<li>Make sure information is easy to find; nothing should be more than 3 clicks away. </li><br />
<li>Avoid using very small fonts and low contrast colors; information should be easy to read. </li><br />
<li>Start documenting your project as early as possible; don’t leave anything to the last minute before the Wiki Freeze. For a complete list of deadlines visit the <a href="">iGEM 2013 calendar</a> </li><br />
<li>Have lots of fun! </li><br />
</ul><br />
</br><br />
</td><br />
</tr><br />
</table></div>
Hliu1423
http://2014.igem.org/Team:Stony_Brook
Team:Stony Brook
2014-06-30T14:08:26Z
<p>Hliu1423: </p>
<hr />
<div>{{CSS/Main}}<br />
<br />
<br />
<html><br />
<br />
<!--main content --><br />
<table width="70%" align="center"><br />
<br />
<br />
<!--welcome box --><br />
<tr><br />
<td style="border:1px solid black;" colspan="3" align="center" height="150px" bgColor=#FF404B><br />
<h1 >WELCOME TO iGEM 2014! </h1><br />
<p>Your team has been approved and you are ready to start the iGEM season!<br />
<br>On this page you can document your project, introduce your team members, document your progress <br> and share your iGEM experience with the rest of the world! </p><br />
<br><br />
<p style="color:#E7E7E7"> <a href="https://2014.igem.org/wiki/index.php?title=Team:Stony_Brook&action=edit"style="color:#FFFFFF"> Click here to edit this page!</a> </p><br />
</td><br />
</tr><br />
<br />
<tr> <td colspan="3" height="5px"> </td></tr><br />
<!-- end welcome box --><br />
<br />
<tr> <br />
<br />
<!--navigation menu --><br />
<td align="center" colspan="3"><br />
<br />
<table width="100%"><br />
<tr heigth="15px"></tr><br />
<tr heigth="75px"> <br />
<br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook"style="color:#000000">Home </a> </td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Team"style="color:#000000"> Team </a> </td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://igem.org/Team.cgi?year=2014&team_name=Stony_Brook"style="color:#000000"> Official Team Profile </a></td><br />
<br />
<td style="border:1px solid black" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Project"style="color:#000000"> Project</a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Parts"style="color:#000000"> Parts</a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Modeling"style="color:#000000"> Modeling</a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Notebook"style="color:#000000"> Notebook</a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Safety"style=" color:#000000"> Safety </a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Attributions"style="color:#000000"> Attributions </a></td><br />
<br />
<br />
<td align ="center"> <a href="https://2014.igem.org/Main_Page"> <img src="https://static.igem.org/mediawiki/igem.org/6/60/Igemlogo_300px.png" width="55px"></a> </td><br />
</tr><br />
</table><br />
<br />
<!--end navigation menu --><br />
</tr><br />
<br />
<br />
</tr><br />
<br />
<br />
<br />
<br />
<br />
</td><br />
<br />
<tr> <td colspan="3" height="15px"> </td></tr><br />
<tr><td bgColor="#e7e7e7" colspan="3" height="1px"> </tr><br />
<tr> <td colspan="3" height="5px"> </td></tr><br />
<br />
<br />
<!--requirements section --><br />
<tr><td colspan="3"> <h3> Requirements </h3></td></tr><br />
<tr><br />
<td width="45%" valign="top"> <br />
<br />
<p> Please be sure to keep these links, your audience will want to find your: </p><br />
<br />
<!-- Links to other team pages --><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook">Home</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Team">Team</a> </li><br />
<li><a href="https://igem.org/Team.cgi?year=2013&team_name=Stony_Brook">Official Team Profile</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Project">Project</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Parts">Parts</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Modeling">Modeling</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Notebook">Notebook</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Safety">Safety</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Attributions">Attributions</a> </li><br />
<br />
</ul><br />
<br />
</td><br />
<br />
<td > </td><br />
<td width="45%"><br />
<br />
<p>There are a few wiki requirements teams must follow:</p><br />
<ul><br />
<li>All pages, images and files must be hosted on the <a href ="https://2014.igem.org/Special:Upload"> 2014.igem.org server</a>. </li><br />
<li>All pages must be created under the team’s name space.</li><br />
<li>As part of your documentation, keep the links from the menu to the left. </li><br />
<li>Do not use flash in wiki code. </li><br />
<li>The <a href="https://static.igem.org/mediawiki/igem.org/6/60/Igemlogo_300px.png"> iGEM logo </a> should be placed on the upper part of every page and should link to <a href="https://2014.igem.org/Main_Page">2014.igem.org</a>.</li><br />
</ul><br />
<p>Visit the <a href="https://2014.igem.org/Wiki_How-To"> Wiki How To page </a> for a complete list of requirements, tips and other useful information. </p><br />
<br />
</td><br />
</tr><br />
<br />
<br />
<tr> <td colspan="3" height="15px"> </td></tr><br />
<tr><td bgColor="#e7e7e7" colspan="3" height="1"> </tr><br />
<br />
<br />
<!--tips --><br />
<tr><td colspan="3" > <h3> Tips </h3></td></tr><br />
<br />
<tr><br />
<td width="45%" valign="top"> <br />
<p>We are currently working on providing teams with some easy to use design templates.<br />
<br> In the meantime you can also view other team wikis for inspiration! Here are some very good examples</p><br />
<br />
<ul><br />
<li> <a href="https://2013.igem.org/Team:SDU-Denmark/"> 2013 SDU Denmark </a> </li><br />
<li> <a href="https://2013.igem.org/Team:SYSU-China">2013 SYSU China</a> </li><br />
<li> <a href="https://2013.igem.org/Team:Shenzhen_BGIC_ATCG"> 2013 Shenxhen BGIG ATCG </a></li><br />
<li> <a href="https://2013.igem.org/Team:Colombia_Uniandes">2013 Colombia Unianades </a></li><br />
<li> <a href="https://2013.igem.org/Team:Lethbridge">2013 Lethbridge</a></li><br />
</ul><br />
<br />
<p>For a full wiki list, you can visit <a href="https://igem.org/Team_Wikis?year=2013">iGEM 2013 web sites </a> and <a href="https://igem.org/Team_Wikis?year=2012">iGEM 2012 web sites</a> lists. </p><br />
</td><br />
<br />
<td> </td><br />
<td width="45%"> <br />
<br />
<p>This wiki will be your team’s first interaction with the rest of the world, so here are a few tips to help you get started: </p><br />
<br />
<ul><br />
<li>State your accomplishments! Tell people what you have achieved from the start. </li><br />
<li>Be clear about what you are doing and what you plan to do.</li><br />
<li>You have a global audience! Consider the different backgrounds that your users come from.</li><br />
<li>Make sure information is easy to find; nothing should be more than 3 clicks away. </li><br />
<li>Avoid using very small fonts and low contrast colors; information should be easy to read. </li><br />
<li>Start documenting your project as early as possible; don’t leave anything to the last minute before the Wiki Freeze. For a complete list of deadlines visit the <a href="">iGEM 2013 calendar</a> </li><br />
<li>Have lots of fun! </li><br />
</ul><br />
</br><br />
</td><br />
</tr><br />
</table></div>
Hliu1423
http://2014.igem.org/Team:Stony_Brook
Team:Stony Brook
2014-06-30T14:07:06Z
<p>Hliu1423: </p>
<hr />
<div>{{CSS/Main}}<br />
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<table width="70%" align="center"><br />
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<td style="border:1px solid black;" colspan="3" align="center" height="150px" bgColor=#FF404B><br />
<h1 >WELCOME TO iGEM 2014! </h1><br />
<p>Your team has been approved and you are ready to start the iGEM season!<br />
<br>On this page you can document your project, introduce your team members, document your progress <br> and share your iGEM experience with the rest of the world! </p><br />
<br><br />
<p style="color:#E7E7E7"> <a href="https://2014.igem.org/wiki/index.php?title=Team:Stony_Brook&action=edit"style="color:#FFFFFF"> Click here to edit this page!</a> </p><br />
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<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook"style="color:#000000">Home </a> </td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Team"style="color:#000000"> Team </a> </td><br />
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<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://igem.org/Team.cgi?year=2014&team_name=Stony_Brook"style="color:#000000"> Official Team Profile </a></td><br />
<br />
<td style="border:1px solid black" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Project"style="color:#000000"> Project</a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Parts"style="color:#000000"> Parts</a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Modeling"style="color:#000000"> Modeling</a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Notebook"style="color:#000000"> Notebook</a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Safety"style=" color:#000000"> Safety </a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Attributions"style="color:#000000"> Attributions </a></td><br />
<br />
<br />
<td align ="center"> <a href="https://2014.igem.org/Main_Page"> <img src="https://static.igem.org/mediawiki/igem.org/6/60/Igemlogo_300px.png" width="55px"></a> </td><br />
</tr><br />
</table><br />
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<!--end navigation menu --><br />
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<tr><td bgColor="#e7e7e7" colspan="3" height="1px"> </tr><br />
<tr> <td colspan="3" height="5px"> </td></tr><br />
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<!--requirements section --><br />
<tr><td colspan="3"> <h3> Requirements </h3></td></tr><br />
<tr><br />
<td width="45%" valign="top"> <br />
<br />
<p> Please be sure to keep these links, your audience will want to find your: </p><br />
<br />
<!-- Links to other team pages --><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook">Home</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Team">Team</a> </li><br />
<li><a href="https://igem.org/Team.cgi?year=2013&team_name=Stony_Brook">Official Team Profile</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Project">Project</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Parts">Parts</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Modeling">Modeling</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Notebook">Notebook</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Safety">Safety</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Attributions">Attributions</a> </li><br />
<br />
</ul><br />
<br />
</td><br />
<br />
<td > </td><br />
<td width="45%"><br />
<br />
<p>There are a few wiki requirements teams must follow:</p><br />
<ul><br />
<li>All pages, images and files must be hosted on the <a href ="https://2014.igem.org/Special:Upload"> 2014.igem.org server</a>. </li><br />
<li>All pages must be created under the team’s name space.</li><br />
<li>As part of your documentation, keep the links from the menu to the left. </li><br />
<li>Do not use flash in wiki code. </li><br />
<li>The <a href="https://static.igem.org/mediawiki/igem.org/6/60/Igemlogo_300px.png"> iGEM logo </a> should be placed on the upper part of every page and should link to <a href="https://2014.igem.org/Main_Page">2014.igem.org</a>.</li><br />
</ul><br />
<p>Visit the <a href="https://2014.igem.org/Wiki_How-To"> Wiki How To page </a> for a complete list of requirements, tips and other useful information. </p><br />
<br />
</td><br />
</tr><br />
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<tr> <td colspan="3" height="15px"> </td></tr><br />
<tr><td bgColor="#e7e7e7" colspan="3" height="1"> </tr><br />
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<!--tips --><br />
<tr><td colspan="3" > <h3> Tips </h3></td></tr><br />
<br />
<tr><br />
<td width="45%" valign="top"> <br />
<p>We are currently working on providing teams with some easy to use design templates.<br />
<br> In the meantime you can also view other team wikis for inspiration! Here are some very good examples</p><br />
<br />
<ul><br />
<li> <a href="https://2013.igem.org/Team:SDU-Denmark/"> 2013 SDU Denmark </a> </li><br />
<li> <a href="https://2013.igem.org/Team:SYSU-China">2013 SYSU China</a> </li><br />
<li> <a href="https://2013.igem.org/Team:Shenzhen_BGIC_ATCG"> 2013 Shenxhen BGIG ATCG </a></li><br />
<li> <a href="https://2013.igem.org/Team:Colombia_Uniandes">2013 Colombia Unianades </a></li><br />
<li> <a href="https://2013.igem.org/Team:Lethbridge">2013 Lethbridge</a></li><br />
</ul><br />
<br />
<p>For a full wiki list, you can visit <a href="https://igem.org/Team_Wikis?year=2013">iGEM 2013 web sites </a> and <a href="https://igem.org/Team_Wikis?year=2012">iGEM 2012 web sites</a> lists. </p><br />
</td><br />
<br />
<td> </td><br />
<td width="45%"> <br />
<br />
<p>This wiki will be your team’s first interaction with the rest of the world, so here are a few tips to help you get started: </p><br />
<br />
<ul><br />
<li>State your accomplishments! Tell people what you have achieved from the start. </li><br />
<li>Be clear about what you are doing and what you plan to do.</li><br />
<li>You have a global audience! Consider the different backgrounds that your users come from.</li><br />
<li>Make sure information is easy to find; nothing should be more than 3 clicks away. </li><br />
<li>Avoid using very small fonts and low contrast colors; information should be easy to read. </li><br />
<li>Start documenting your project as early as possible; don’t leave anything to the last minute before the Wiki Freeze. For a complete list of deadlines visit the <a href="">iGEM 2013 calendar</a> </li><br />
<li>Have lots of fun! </li><br />
</ul><br />
</br><br />
</td><br />
</tr><br />
</table></div>
Hliu1423
http://2014.igem.org/Team:Stony_Brook
Team:Stony Brook
2014-06-30T14:06:35Z
<p>Hliu1423: </p>
<hr />
<div>{{CSS/Main}}<br />
<br />
<br />
<html><br />
<br />
<br />
<tr> <br />
<br />
<!--navigation menu --><br />
<td align="center" colspan="3"><br />
<br />
<table width="100%"><br />
<tr heigth="15px"></tr><br />
<tr heigth="75px"> <br />
<br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook"style="color:#000000">Home </a> </td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Team"style="color:#000000"> Team </a> </td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://igem.org/Team.cgi?year=2014&team_name=Stony_Brook"style="color:#000000"> Official Team Profile </a></td><br />
<br />
<td style="border:1px solid black" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Project"style="color:#000000"> Project</a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Parts"style="color:#000000"> Parts</a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Modeling"style="color:#000000"> Modeling</a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Notebook"style="color:#000000"> Notebook</a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Safety"style=" color:#000000"> Safety </a></td><br />
<br />
<td style="border:1px solid black;" align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7> <br />
<a href="https://2014.igem.org/Team:Stony_Brook/Attributions"style="color:#000000"> Attributions </a></td><br />
<br />
<br />
<td align ="center"> <a href="https://2014.igem.org/Main_Page"> <img src="https://static.igem.org/mediawiki/igem.org/6/60/Igemlogo_300px.png" width="55px"></a> </td><br />
</tr><br />
</table><br />
<br />
<!--end navigation menu --><br />
</tr><br />
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</tr><br />
<br />
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</td><br />
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<tr> <td colspan="3" height="15px"> </td></tr><br />
<tr><td bgColor="#e7e7e7" colspan="3" height="1px"> </tr><br />
<tr> <td colspan="3" height="5px"> </td></tr><br />
<br />
<br />
<!--requirements section --><br />
<tr><td colspan="3"> <h3> Requirements </h3></td></tr><br />
<tr><br />
<td width="45%" valign="top"> <br />
<br />
<p> Please be sure to keep these links, your audience will want to find your: </p><br />
<br />
<!-- Links to other team pages --><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook">Home</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Team">Team</a> </li><br />
<li><a href="https://igem.org/Team.cgi?year=2013&team_name=Stony_Brook">Official Team Profile</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Project">Project</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Parts">Parts</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Modeling">Modeling</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Notebook">Notebook</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Safety">Safety</a> </li><br />
<li><a href="https://2014.igem.org/Team:Stony_Brook/Attributions">Attributions</a> </li><br />
<br />
</ul><br />
<br />
</td><br />
<br />
<td > </td><br />
<td width="45%"><br />
<br />
<p>There are a few wiki requirements teams must follow:</p><br />
<ul><br />
<li>All pages, images and files must be hosted on the <a href ="https://2014.igem.org/Special:Upload"> 2014.igem.org server</a>. </li><br />
<li>All pages must be created under the team’s name space.</li><br />
<li>As part of your documentation, keep the links from the menu to the left. </li><br />
<li>Do not use flash in wiki code. </li><br />
<li>The <a href="https://static.igem.org/mediawiki/igem.org/6/60/Igemlogo_300px.png"> iGEM logo </a> should be placed on the upper part of every page and should link to <a href="https://2014.igem.org/Main_Page">2014.igem.org</a>.</li><br />
</ul><br />
<p>Visit the <a href="https://2014.igem.org/Wiki_How-To"> Wiki How To page </a> for a complete list of requirements, tips and other useful information. </p><br />
<br />
</td><br />
</tr><br />
<br />
<br />
<tr> <td colspan="3" height="15px"> </td></tr><br />
<tr><td bgColor="#e7e7e7" colspan="3" height="1"> </tr><br />
<br />
<br />
<!--tips --><br />
<tr><td colspan="3" > <h3> Tips </h3></td></tr><br />
<br />
<tr><br />
<td width="45%" valign="top"> <br />
<p>We are currently working on providing teams with some easy to use design templates.<br />
<br> In the meantime you can also view other team wikis for inspiration! Here are some very good examples</p><br />
<br />
<ul><br />
<li> <a href="https://2013.igem.org/Team:SDU-Denmark/"> 2013 SDU Denmark </a> </li><br />
<li> <a href="https://2013.igem.org/Team:SYSU-China">2013 SYSU China</a> </li><br />
<li> <a href="https://2013.igem.org/Team:Shenzhen_BGIC_ATCG"> 2013 Shenxhen BGIG ATCG </a></li><br />
<li> <a href="https://2013.igem.org/Team:Colombia_Uniandes">2013 Colombia Unianades </a></li><br />
<li> <a href="https://2013.igem.org/Team:Lethbridge">2013 Lethbridge</a></li><br />
</ul><br />
<br />
<p>For a full wiki list, you can visit <a href="https://igem.org/Team_Wikis?year=2013">iGEM 2013 web sites </a> and <a href="https://igem.org/Team_Wikis?year=2012">iGEM 2012 web sites</a> lists. </p><br />
</td><br />
<br />
<td> </td><br />
<td width="45%"> <br />
<br />
<p>This wiki will be your team’s first interaction with the rest of the world, so here are a few tips to help you get started: </p><br />
<br />
<ul><br />
<li>State your accomplishments! Tell people what you have achieved from the start. </li><br />
<li>Be clear about what you are doing and what you plan to do.</li><br />
<li>You have a global audience! Consider the different backgrounds that your users come from.</li><br />
<li>Make sure information is easy to find; nothing should be more than 3 clicks away. </li><br />
<li>Avoid using very small fonts and low contrast colors; information should be easy to read. </li><br />
<li>Start documenting your project as early as possible; don’t leave anything to the last minute before the Wiki Freeze. For a complete list of deadlines visit the <a href="">iGEM 2013 calendar</a> </li><br />
<li>Have lots of fun! </li><br />
</ul><br />
</br><br />
</td><br />
</tr><br />
</table></div>
Hliu1423