Team:Berkeley

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  <div class="col6" ><img src="https://static.igem.org/mediawiki/2011/1/12/ProjectSummaryHeader.jpg" width="480"> <p> Biosensors have widespread applications ranging from diagnostics to environmental monitoring. Vibrio cholerae's ToxR system can be used as a component in biological devices capable of detecting a wide variety of molecules. A periplasmic domain causes ToxR homodimerization, activating transcription of the ctx promoter. By replacing the periplasmic domain of ToxR with existing or engineered ligand-dependent homodimers, we hope to link ToxR dimerization (and gene expression) to the presence of specific ligands. Initially, ToxR constructs proved to be toxic to E. coli. To address ToxR toxicity, we screened microarray data for promoters that exhibited stress-based down regulation. We constructed a negative feedback system with the rffGH promoter, which permits the use of potentially toxic proteins like our various ToxR chimeras. By fusing existing or engineered ligand dependent homodimers to ToxR, this modular system can be applied to develop new biosensors. </p>
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<p>Biosensors have widespread applications ranging from diagnostics to environmental monitoring. Vibrio cholerae's ToxR system can be used as a component in biological sensing devices. ToxS causes ToxR homodimerization, activating transcription of the ctx promoter. By replacing the periplasmic domain of ToxR with existing or engineered ligand-dependent homodimers, we hope to link ToxR dimerization (and gene expression) to the presence of specific ligands. Initially, ToxR constructs proved toxic to E. coli. We built a stress-regulated transcription system that drives relatively high expression of toxic proteins. This allowed us to further engineer ToxR chimeras. We fused an estrogen-dependent dimer with ToxR hoping to create an estrogen biosensor. We observed a range of constitutive phenotypes and plan more experiments to engineer a dose-dependent transcriptional response to estrogen. By fusing existing or engineered ligand dependent homodimers to ToxR, this modular system can be used to build new biosensors. </p>
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<div class="col3"><a href="https://2011.igem.org/Team:Berkeley/Project#ToxR"> <img src="https://static.igem.org/mediawiki/igem.org/1/10/Subtitlepic1.jpg"  
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<div class="col3-2"style="background-color:#101a4d;"><a href="https://2011.igem.org/Team:Berkeley/Project#ToxR"> <img src="https://static.igem.org/mediawiki/igem.org/1/10/Subtitlepic1.jpg"  
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  <p> A protein with great potential as a general biosensor system.</p> </div>
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  <p style="text-align:center; color:#CECECE;"> A protein with great potential as a general biosensor system.</p> </div>
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<p> Our method for expressing interesting (but toxic) proteins.</p> </div>
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<p style="text-align:center; color:#CECECE;"> Chimeric proteins that drive translation off of the Pctx promoter.</p></div>
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<div class="col3"><a href="https://2011.igem.org/Team:Berkeley/Project#ToxRChimera"><img src="https://static.igem.org/mediawiki/igem.org/f/fd/Subtitlepic3.jpg"  
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<p> Chimeric proteins that drive translation off of the Pctx promoter.</p></div>
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<p style="text-align:center; color:#CECECE;"> Our method for expressing interesting (but toxic) proteins.</p> </div>
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<div class="col3"><a href="https://2011.igem.org/Team:Berkeley/Project#EstrogenReceptor"><img src="https://static.igem.org/mediawiki/2011/c/c8/Subtitlepic4.jpg"  
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  <p> Bacteria designed to detect environmental estrogen contamination.</p> </div>
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  <p style="text-align:center; color:#CECECE;"> Bacteria engineered to detect the presence of estrogen. <br></p></div>
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<p>We are a cohesive team of 7 undergraduates and 3 advisers. iGEM has taught an incredible amount of information about synthetic biology and ourselves. We have learned many different techniques in the lab, but the most important part about iGEM is that we learned to be critical thinkers. We created a project together, carried it out together, and we are going to present at the Jamboree together. Of course this was no easy task and took us twice as long as it would have taken a graduate student, but when we get it right, it is because we figured out how to do it on our own. This accomplishment increases confidence. In a few months, our team has worked together effectively to get a complicated project done. This means we have to work tightly together and it forces us to trust one another. </p>
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<p>We are Team Berkeley, a cohesive unit of 7 undergraduates and 3 advisers. Earlier this year we planned a complex and risky project, given the short amount of time iGEM made available. We quickly learned each others strengths and weaknesses and developed standard systems of organizational management in order to synchronize our efforts for the many parallel tasks at hand. We created protocols, shared them with one another, and worked together on troubleshooting. Using google docs to keep up with the status of cloning projects, the results of assays, material logistics, or the final steps required to complete a project ensured that through the months of hard work, we fine-tuned our ability to work together. As a team, we have learned firsthand how the synthetic biology community relies on the goal-oriented cooperation of skilled individuals from very different backgrounds and with very different skill sets. Some of us have strong engineering backgrounds while others of us have strong biology backgrounds, but we all share a desire to build synthetic biological systems that solve human problems. We are proud of the project that we have created which we will present at the Jamboree together in October.</p>
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<p> <b> The UC Berkeley iGEM team would like to thank Agilent for their financial support and Synberc, for their administrative support. </b></p>
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<p style="text-align:left; color:#CECECE; font-size:13px; font-weight:bold; padding:3px"> The UC Berkeley iGEM team would like to thank Autodesk and Agilent for their financial support and Synberc, for their administrative support. </p>
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Latest revision as of 03:39, 29 September 2011

Berkeley iGEM 2011

header
Mercury

Biosensors have widespread applications ranging from diagnostics to environmental monitoring. Vibrio cholerae's ToxR system can be used as a component in biological sensing devices. ToxS causes ToxR homodimerization, activating transcription of the ctx promoter. By replacing the periplasmic domain of ToxR with existing or engineered ligand-dependent homodimers, we hope to link ToxR dimerization (and gene expression) to the presence of specific ligands. Initially, ToxR constructs proved toxic to E. coli. We built a stress-regulated transcription system that drives relatively high expression of toxic proteins. This allowed us to further engineer ToxR chimeras. We fused an estrogen-dependent dimer with ToxR hoping to create an estrogen biosensor. We observed a range of constitutive phenotypes and plan more experiments to engineer a dose-dependent transcriptional response to estrogen. By fusing existing or engineered ligand dependent homodimers to ToxR, this modular system can be used to build new biosensors.

A protein with great potential as a general biosensor system.

Chimeric proteins that drive translation off of the Pctx promoter.

Our method for expressing interesting (but toxic) proteins.

Bacteria engineered to detect the presence of estrogen.


We are Team Berkeley, a cohesive unit of 7 undergraduates and 3 advisers. Earlier this year we planned a complex and risky project, given the short amount of time iGEM made available. We quickly learned each others strengths and weaknesses and developed standard systems of organizational management in order to synchronize our efforts for the many parallel tasks at hand. We created protocols, shared them with one another, and worked together on troubleshooting. Using google docs to keep up with the status of cloning projects, the results of assays, material logistics, or the final steps required to complete a project ensured that through the months of hard work, we fine-tuned our ability to work together. As a team, we have learned firsthand how the synthetic biology community relies on the goal-oriented cooperation of skilled individuals from very different backgrounds and with very different skill sets. Some of us have strong engineering backgrounds while others of us have strong biology backgrounds, but we all share a desire to build synthetic biological systems that solve human problems. We are proud of the project that we have created which we will present at the Jamboree together in October.


The UC Berkeley iGEM team would like to thank Autodesk and Agilent for their financial support and Synberc, for their administrative support.


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