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Team:British Columbia/Data
From 2011.igem.org
Contents |
Our System
Data for our Favourite New Parts
GAL galactose-inducible yeast promoter
Promoter can be induced to express a marker gene (GFP) when exposed to galactose in the absence of glucose.
This promoter was taken from the S. cerevisiae Advanced Gateway Destination Vectors.
GPD constitutive yeast promoter
Promoter for constitutively high expression.
This promoter was taken from the S. cerevisiae Advanced Gateway Destination Vectors.
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.
Cineole Synthase
Alpha-Pinene Synthase
Beta-Pinene Synthase
ERG20
The ERG20 gene is an essential gene naturally found in yeast. It encodes for an enzyme that assists the mevalonate pathway to produce FPP and GPP from I-PP and DMA-PP. Our iGEM team would like to produce more GPP than FPP because GPP is a precursor in producing monoterpenes where as FPP produces diterpenoids instead.
The iGEM team aims to characterize this gene in vivo in S. cerevisiae yeast. The products wil be sent for GC-MS. Furthermore, this will be made into a biobrick and become an addition to the iGEM parts registry.
Reference: Ignea C., Cvetkovic I., Loupassaki S., Kefalas P., Johnson C. B, Kampranis S. C, Makris A. M. Improving yeast strains using recyclable integration cassettes, for the production of plant terpenoids. 2011 10:4
Reference: Norihiko Misawa. Pathway engineering for functional isoprenoids. 2011.01.002 22:1-7
erg20-2
The erg20-2 gene is a mutant of the ERG20 gene. It is a mutant because of the K197 to E substitution. It also for an enzyme that assists the mevalonate pathway to produce FPP and GPP from I-PP and DMA-PP. It was observed that this gene produces "25% FPP and 75% GPP instead of 75% FPP and 25% GPP" when compared with the wild type ERG20 gene.
The iGEM team aims to characterize this gene in vivo in S. cerevisiae yeast. The products wil be sent for GC-MS. Furthermore, this will be made into a biobrick and become an addition to the iGEM parts registry. In particular, we would like to determine whether more monoterpenes are produced with this mutant gene than the wild type ERG20 gene.
Reference: Oswald M., Fischer M., Dirninger N., Karst F. Monoterpenoid biosynthesis in Saccharomyces cerevisiae. 2007 413:421
IDI1
HMG2
Data for Pre-Existing Parts
BBa_K118025: Limonene Synthase Composite + Lac promotor BBa_K118024: Limonene Synthase Composite BBa_J63006: Yeast GAL1 Promoter BBa_K115056: E. coli Isopentenyl Diphosphate Isomerase
