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Team:British Columbia/Data
From 2012.igem.org
Contents |
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.
