Team:Wisconsin-Madison

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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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<h1>Introduction</h1>
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<p1> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;With Badger football season upon us, close to 10,000 Badger fans made their way to Camp Randall Stadium on Sunday, Aug. 19 to get up-close and personal with Wisconsin players and coaches at the annual Family Fun Day. It was a day for autographs, face painting and football fun.</p1>
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<p2>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;At left: Wisconsin freshman quarterback Chase Knox (9) plays defense as Luke Brown, 6, runs the ball at mid-field during the free event.&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;With Badger football season upon us, close to 10,000 Badger fans made their way to Camp Randall Stadium on Sunday, Aug. 19 to get up-close and personal with Wisconsin players and coaches at the annual Family Fun Day. It was a day for autographs, face painting and football fun.</p2>
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<a href="https://2012.igem.org/Team:Wisconsin-Madison/team#member">Members</a>
 
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<a href="https://2012.igem.org/Team:Wisconsin-Madison/team#advisor">Advisors</a>
 
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<a href="https://2012.igem.org/Team:Wisconsin-Madison/team#contribution&attribution">Contributions &amp; Attributions</a>
 
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<a href="https://2012.igem.org/Team:Wisconsin-Madison/project/limonene">Limonene production in E.coli</a>
 
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<a href="https://2012.igem.org/Team:Wisconsin-Madison/parts/???">Part two</a>
 
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<a href="https://2012.igem.org/Team:Wisconsin-Madison/parts/???">Part three</a>
 
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<a href="https://2012.igem.org/Team:Wisconsin-Madison/parts/???">Part four</a>
 
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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.









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