Team:Wisconsin-Madison

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<a href="https://2011.igem.org/Team:Wisconsin-Madison">Main</a>
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<a href="https://2011.igem.org/Team:Wisconsin-Madison/whatisigem">What is iGEM?</a>
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<a href="https://2011.igem.org/Team:Wisconsin-Madison/ca">Contributions &amp; Attributions</a>
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<a href="https://2011.igem.org/Team:Wisconsin-Madison/projectoverview">Overview</a>
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<a href="https://2011.igem.org/Team:Wisconsin-Madison/ethanol">Ethanol Sensor</a>
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<a href="https://2011.igem.org/Team:Wisconsin-Madison/alkane">Alkane Sensor</a>
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                <a href="https://2011.igem.org/Team:Wisconsin-Madison/directedevolution">Directed Evolution</a>
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<a href="https://2011.igem.org/Team:Wisconsin-Madison/bmc">Microcompartment</a>
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                <a href="https://2011.igem.org/Team:Wisconsin-Madison/parts">Parts</a>
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                <a href="https://2011.igem.org/Team:Wisconsin-Madison/teamoverview">Overview</a>
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<a href="https://2011.igem.org/Team:Wisconsin-Madison/teammembers">Members</a>
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<a href="https://2011.igem.org/Team:Wisconsin-Madison/teamadvisors">Advisors</a>
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<a href="https://2011.igem.org/Team:Wisconsin-Madison/scienceolympiad">Science Olympiad</a>
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<p><strong><a href="https://2011.igem.org/Main_Page">iGEM 2011</a>: Wisconsin-Madison</strong><p>
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This summer, the UW-Madison <a href="https://2011.igem.org/Team:Wisconsin-Madison/whatisigem">iGEM</a> team will be working on streamlining the <a href="https://2011.igem.org/Team:Wisconsin-Madison/biofuel">biofuel</a> discovery and production processes through the use of <a href="https://2011.igem.org/Team:Wisconsin-Madison/biosensor">biosensors</a>. The necessity for sustainable, economical sources of fuel is ever growing, and <a href="http://www.wisc.edu/">UW-Madison</a> is a forefront institution in the hunt for such supplies. In association with the Great Lakes Bioenergy Research Center (<a href="http://www.glbrc.org/">GLBRC</a>), we are creating new E. coli biosensors that can accelerate high throughput screening of potential fuel sources. We’re specifically interested in <a href="https://2011.igem.org/Team:Wisconsin-Madison/ethanol">ethanol</a> and <a href="https://2011.igem.org/Team:Wisconsin-Madison/alkane">alkane</a>, derived from sources ranging from cellulose to metabolically engineered E. coli.
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We have found regulatory systems which respond to each of the biofuels of interest, and are using standard <a href="https://2011.igem.org/Team:Wisconsin-Madison/biobrick">BioBricks</a> assembly to create E. coli strains which can be used to perform <a href="https://2011.igem.org/Team:Wisconsin-Madison/platereader"> fluorescence-based</a> assays. By using fluorescent biosensors, we hope to lower costs (in both equipment and cost-per-sample) while maintaining a high degree of accuracy. In the interest of creating robust and accurate assays, we are also attempting to increase the magnitude and range of the linear fluorescence response through directed evolution. We hope to leverage multiple selections to both decrease basal fluorescence and increase the point where the response becomes saturated.
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<p>
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As a more direct approach to increasing microbial biofuel yields, we also pursued the use of <a href="https://2011.igem.org/Team:Wisconsin-Madison/bmc">bacterial microcompartments</a> (BMCs) as scaffolding for key enzymes in the ethanol producing and sensing processes. Through localizing crucial anabolic enzymes, as well as the beginning of our sensing cascades, to the BMC surfaces, we can increase fuel titers as well as our reliability in accurately sensing them. However, due to the complexity of this project and the time investment needed, we have stopped work on the BMC. More information can be found on the <a href="https://2011.igem.org/Team:Wisconsin-Madison/bmc">Microcompartments</a> page.
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We hope that when combined our projects will produce a useful toolkit for researchers in and beyond the GLBRC to aid the discovery and engineering of promising fuel sources and processes.
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<p><br> <p>
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<b>Click <a href="https://2011.igem.org/Team:Wisconsin-Madison/ca">here</a> to see our contributions and attributions page.</b>
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<b>And <a href="https://2011.igem.org/Team:Wisconsin-Madison/data">here</a> to see our data page.</b>
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      <p class = "classtheoverview"> <strong> The Translational Coupling Cassette: a tool for evaluating the translation of heterologous proteins in <i>Escherichia coli</i>. </strong></p>
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      <p align="left" class = "classtheinlinecontent2"> A powerful method for the production of novel metabolites is the expression of heterologous enzymes in a bacterial host. A common challenge when using non-native genes in metabolic engineering is determining if they are being properly expressed. To address this issue, we have constructed a BioFusion-compatible system for testing the translation of a gene of interest. This system couples the translation of the target gene to a fluorescent reporter gene; fluorescence will only be detected when the target gene is entirely translated. This construct enables synthetic biologists to quickly determine if a gene is being expressed without the need for costly antibodies or analytical instruments (e.g. mass spectrometry). Currently, we are utilizing this cassette to optimize the expression of limonene synthase, an enzyme that catalyzes the production of limonene, a monoterpene with potential as a renewable jet fuel.</p>
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Latest revision as of 20:08, 26 October 2012


The Translational Coupling Cassette: a tool for evaluating the translation of heterologous proteins in Escherichia coli.

A powerful method for the production of novel metabolites is the expression of heterologous enzymes in a bacterial host. A common challenge when using non-native genes in metabolic engineering is determining if they are being properly expressed. To address this issue, we have constructed a BioFusion-compatible system for testing the translation of a gene of interest. This system couples the translation of the target gene to a fluorescent reporter gene; fluorescence will only be detected when the target gene is entirely translated. This construct enables synthetic biologists to quickly determine if a gene is being expressed without the need for costly antibodies or analytical instruments (e.g. mass spectrometry). Currently, we are utilizing this cassette to optimize the expression of limonene synthase, an enzyme that catalyzes the production of limonene, a monoterpene with potential as a renewable jet fuel.









Retrieved from "http://2012.igem.org/Team:Wisconsin-Madison"