Team:Wisconsin-Madison

From 2012.igem.org

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       <p align="left" class="classtheinlinecontent"> <strong style="font-style:italic; color: rgb(183, 1, 1);">TCC</strong>
       <p align="left" class="classtheinlinecontent"> <strong style="font-style:italic; color: rgb(183, 1, 1);">TCC</strong>
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       <p align="left" class="classtheinlinecontent">Limonene is a colorless, liquid hydrocarbon that smells like lemon. It is found in the oils of citrus fruits and is primarily used as a cleaning agent, solvent, and flavorful food additive. Limonene also possesses chemical properties that make it an ideal jet fuel due to it's low freezing point and high combustibility. E. coli already has part of the metabolic pathway required to produce limonene, however we will be inserting genes from S. cerevisiae to create the mevalonate pathway in the cell. We will also codon optimize the Limonene Synthase (LimS1) gene which is the final step in converting GPP to limonene. Gas chromatography/mass spectrometry will be used to determine how much limonene the E. coli is producing. </p>
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       <p align="left" class="classtheinlinecontent">In synthetic biology, one of the most powerful tools a research has is the ability to express heterologous genes in Escherichia coli. Unfortunately this does not always work. 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 diagnosing the failure of a gene expression at the translational level. 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 antibiodies or analytical instruments.</p>
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<p align="left" class="classtheinlinecontent" style="font-style:italic">This tool is based on a phenomenon known as translational coupling. </p>
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Revision as of 21:17, 21 September 2012


Engineering the limonene production metabolic pathway: TCC.

Creating a tool to evaluate the translation of heterologous genes in Eschericia coli.



TCC

In synthetic biology, one of the most powerful tools a research has is the ability to express heterologous genes in Escherichia coli. Unfortunately this does not always work. 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 diagnosing the failure of a gene expression at the translational level. 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 antibiodies or analytical instruments.

This tool is based on a phenomenon known as translational coupling.






Translational couplding cassette


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