Team:British Columbia/Characterized

From 2011.igem.org

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Latest revision as of 15:22, 26 August 2011

Team: British Columbia - 2011.igem.org

Here are the new parts we aim to characterize:

A variety of monoterpene synthases that produce alpha- and beta- pinene, carene and cineole.

ERG20 and erg20-2

IDI1

HMG20

GAL galactose-inducible yeast promoter

GPD constitutive yeast promoter

3-Carene Synthase

The 3-carene synthase is a monoterpene synthase isolated from Picea sitchensis by Chris Keeling and Dr. Joerg Bohlmann. This 3-carene synthase was found to confer resistance to the white pine weevil, part of the reason why the iGEM team decided to characterize this part in yeast. This synthase converts farnesyl pyrophosphate to 66.4% (+)-3-Carene, 16.3% Terpinolene, 2.7% (-)-alpha-pinene, 2.5% Terpinen-4-ol, 2.1% (-)-beta-phellandrene, 2.1% myrcene, 1.4% alpha-Terpinol, 0.8% gamma-terpinene, 1% others.

The 3-carene 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.

The iGEM team aims to characterize the 3-carene synthase in vivo in S. cerevisiae yeast. The products wil be sent for GC-MS.

Reference: Keeling, CI, Weisshar S, Ralph SG, Jancsik S, Hamberger B, Dullat HK, and Joerg Bohlmann. Transcriptome mining, functional characterization, and phylogeny of a large terpene synthase gene family in spruce (Picea spp.) BMC. 2011 March 7 11:43.

Here are the existing parts we aim to characterize/improve:

BBa_K118025: Limonene Synthase Composite + Lac promotor
BBa_K118024: Limonene Synthase Composite
BBa_J63006: Yeast GAL1 Promoter

@@ Line 19: / Line 19: @@
 ===3-Carene Synthase===
-NCBI: HQ426167
-The 3-carene synthase is a monoterpene synthase isolated from Picea sitchensis by Chris Keeling and Dr. Joerg Bohlmann. This 3-carene synthase was found to be resistant to white pine weevil, part of the reason why the iGEM team decided to characterize this part in yeast (2011 Keeling). This synthase converts farnesyl pyrophosphate to 66.4% (+)-3-Carene, 16.3% Terpinolene, 2.7% (-)-alpha-pinene, 2.5% Terpinen-4-ol, 2.1% (-)-beta-phellandrene, 2.1% myrcene, 1.4% alpha-Terpinol, 0.8% gamma-terpinene, 1% others (2011 Keeling).
+The 3-carene synthase is a monoterpene synthase isolated from Picea sitchensis by Chris Keeling and Dr. Joerg Bohlmann. This 3-carene synthase was found to confer resistance to the white pine weevil, part of the reason why the iGEM team decided to characterize this part in yeast. This synthase converts farnesyl pyrophosphate to 66.4% (+)-3-Carene, 16.3% Terpinolene, 2.7% (-)-alpha-pinene, 2.5% Terpinen-4-ol, 2.1% (-)-beta-phellandrene, 2.1% myrcene, 1.4% alpha-Terpinol, 0.8% gamma-terpinene, 1% others.
-The 3-carene synthase has been previously characterized by expression in C41 DE3 cells, with a His-tag in vitro purification. The purified enzymes were assayed with GPP and sent for GCMS (2011 Keeling).
-The iGEM team aims to characterize the 3-carene synthase in vivo in sacchromeyces cerevisae. The products wil be sent for GCMS.
-Sources:
-(1) Keeling, CI, Weisshar S, Ralph SG, Jancsik S, Hamberger B, Dullat HK, and Joerg Bohlmann. Transcriptome mining, functional characterization, and phylogeny of a large terpene synthase gene family in spruce (Picea spp.) 2011. BMC. 11: 43
+The 3-carene 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.
+The iGEM team aims to characterize the 3-carene synthase in vivo in ''S. cerevisiae'' yeast. The products wil be sent for GC-MS.
+Reference: Keeling, CI, Weisshar S, Ralph SG, Jancsik S, Hamberger B, Dullat HK, and Joerg Bohlmann. Transcriptome mining, functional characterization, and phylogeny of a large terpene synthase gene family in spruce (Picea spp.) BMC. 2011 March 7 11:43.
 ==Here are the existing parts we aim to characterize/improve:==