Team:British Columbia/Data

From 2012.igem.org

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'''Our System'''
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<b>Our System</b></br></br>
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'''Data for our Favourite New Parts'''
 
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<html><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K517002"><b>Main Page</a> - Alpha-Pinene Synthase Gene, BBa_K517002:</b> The (-)-a-pinene synthase (PsTPS-Pin) was isolated from Sitka spruce and converts geranyl pyrophosphate to 83.4%(-)-alpha-pinene, 12.6%(-)-beta-pinene, 2.1% Linalool, 1.0% (-)-beta-phellandrene, 0.4% Camphene, and 0.4% myrcene. We characterized it by expression in C41 DE3 E. coli followed by a His-tag purification. The purified enzymes were assayed in vitro with GPP and sent for GC-MS, which confirmed alpha- and beta-pinene as products. </br></br>
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<b>Data for our Favourite New Parts</b></br></br>
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<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K517002"><b>Main Page</a> - Alpha-Pinene Synthase Gene, BBa_K517002:</b> The (-)-a-pinene synthase (PsTPS-Pin) was isolated from Sitka spruce and converts geranyl pyrophosphate to 83.4%(-)-alpha-pinene, 12.6%(-)-beta-pinene, 2.1% Linalool, 1.0% (-)-beta-phellandrene, 0.4% Camphene, and 0.4% myrcene. We characterized it by expression in C41 DE3 E. coli followed by a His-tag purification. The purified enzymes were assayed in vitro with GPP and sent for GC-MS, which confirmed alpha- and beta-pinene as products. </br></br>
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'''Data for Pre-Existing Parts'''
 
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<b>Data for Pre-Existing Parts</b></br></br>
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<b><a href="http://partsregistry.org/Part:BBa_K118025:Experience">Experience</a> - Limonene Synthase Composite + Lac promotor, BBa_K118025 (Edinburgh, iGEM 2008):</b> This part was previously uncharacterized. We expressed it in C41 <i>E. coli</i> that possesses a pRARE plasmid that enables it to translate proteins with rare codons. The limonene synthase was purified from a culture and subjected to an enzymatic assay for its ability to synthesize limonene monoterpene from GPP substrate. GC-MS analysis showed that limonene was produced, demonstrating that this part works!</br>
<b><a href="http://partsregistry.org/Part:BBa_K118025:Experience">Experience</a> - Limonene Synthase Composite + Lac promotor, BBa_K118025 (Edinburgh, iGEM 2008):</b> This part was previously uncharacterized. We expressed it in C41 <i>E. coli</i> that possesses a pRARE plasmid that enables it to translate proteins with rare codons. The limonene synthase was purified from a culture and subjected to an enzymatic assay for its ability to synthesize limonene monoterpene from GPP substrate. GC-MS analysis showed that limonene was produced, demonstrating that this part works!</br>
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'''We've also characterized the following parts'''
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<b>We've also characterized the following parts</b></br></br>
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<html><a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K517001"><b>Main Page</a> - GPD constitutive yeast promoter, BBa_K517001:</b> Promoter for constitutively high expression. We characterized this part by placing a reporter GFP gene downstream of the promoter and determined that GFP was expressed regardless of whether the yeast was in glucose, raffinose or galactose media.</html>
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<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K517001"><b>Main Page</a> - GPD constitutive yeast promoter, BBa_K517001:</b> Promoter for constitutively high expression. We characterized this part by placing a reporter GFP gene downstream of the promoter and determined that GFP was expressed regardless of whether the yeast was in glucose, raffinose or galactose media.</html>

Revision as of 22:09, 29 September 2012

British Columbia - 2012.igem.org Our System

Data for our Favourite New Parts

Main Page - Alpha-Pinene Synthase Gene, BBa_K517002: The (-)-a-pinene synthase (PsTPS-Pin) was isolated from Sitka spruce and converts geranyl pyrophosphate to 83.4%(-)-alpha-pinene, 12.6%(-)-beta-pinene, 2.1% Linalool, 1.0% (-)-beta-phellandrene, 0.4% Camphene, and 0.4% myrcene. We characterized it by expression in C41 DE3 E. coli followed by a His-tag purification. The purified enzymes were assayed in vitro with GPP and sent for GC-MS, which confirmed alpha- and beta-pinene as products.

Data for Pre-Existing Parts

Experience - Limonene Synthase Composite + Lac promotor, BBa_K118025 (Edinburgh, iGEM 2008): This part was previously uncharacterized. We expressed it in C41 E. coli that possesses a pRARE plasmid that enables it to translate proteins with rare codons. The limonene synthase was purified from a culture and subjected to an enzymatic assay for its ability to synthesize limonene monoterpene from GPP substrate. GC-MS analysis showed that limonene was produced, demonstrating that this part works!

We've also characterized the following parts

Main Page - GPD constitutive yeast promoter, BBa_K517001: Promoter for constitutively high expression. We characterized this part by placing a reporter GFP gene downstream of the promoter and determined that GFP was expressed regardless of whether the yeast was in glucose, raffinose or galactose media.

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