Team:British Columbia/Accomplishments

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Team: British Columbia - 2011.igem.org

Project Accomplishments

Contents

Wet Laboratory Results

<h2>1. In vitro assay production of monoterpene in bacteria

GPP Assay

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Modelling Results

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.

Main Page - Beta-Pinene Synthase Gene, BBa_K517003: The (-)-b-pinene synthase (PsTPS-Pin2) synthase is a monoterpene synthase isolated from Sitka spruce converts geranyl pyrophosphate to 70.5%(-)-beta-pinene, 29.5%(-)-alpha-pinene. (-)-b-pinene synthase has been previously characterized 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.

Main Page - GAL galactose-inducible yeast promoter, BBa_K517000: Promoter can be induced when exposed to galactose in the absence of glucose. We characterized this part by placing a reporter GFP gene downstream of the promoter and determined that GFP expression was up-regulated when the yeast was shifted to galactose media.

Human Practice



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.