Revision as of 15:32, 25 September 2012

Constitutive Promoters

Responsible: Georg Schützinger

For the expression of of proteins like limonene, thaumatin and others promoters are essential.

Background and principles

Jie Sun et al. describes 14 constitutive promoters from Saccharomyces cerevisiae, and its reaction at different culture conditions, oxy(-)/glu(+),oxy(+)/glu(+). 2 Where described as higher strength promoters TEF1p, TPI1p, and 7 as medium strenth pr. GPM1p, GPDp (TDH3p), FBA1p, PDC1p, ENO2p, PYK1p, and TEF2p. With exception to the strongest TEF1p all exhibit nearly the same expression values with varying oxygen conditions. TEF1p was also tested for small glucose concentrations showed 100% expression levels with oxygen limitation and 70 % without oxygenlimitation after 80 hours of cell cultivation.

A.Blount et al. states that repeated use of the same biological parts is not possible since the cells recombine long stretches of homologous DNA

Siavash Partow et al. compares several constitutive Promoters during varying Glucose concentrations Media:Tum12Promoterstärken.pdf, TEF1, HXT7,PGK1 are shown to be the most suitable ones. They also show the fusion of two promoters for bidirectional control of two genes namely TEF1 and PGK1 with comparable activity in both states Media:Tum12Promoterfusion.pdf, two constructs were successfully used pSP-G1-Vector and pSP-G2.

Media:Tum12Fusionspromoter_Vector.pdf

Elke Nevoigt et al. generated a library of TEF1 promotermutants for finetuning of Expression Media:Tum12 TEF-Promoter-mutants.pdf

General remarks and issues

TEF1p guarantues relative strong expression values under varying conditions as it is the case for brewing.

Results

Gel picture of finished Constructs

Results of luciferase assay

References

Media:Tum12_Characterization_of_different_promoters_for_designing_a_new_expression_vector_in_Saccharomyces_cerevisiae.pdf‎

Media:Tum12_Rational_Diversification_of_a_Promoter_Providing_Fine-_Tuned_Expression_and_Orthogonal_Regulation_for_Synthetic_Biology.pdf‎

Media:Tum12 Cloning and Characterization of a Panel of Constitutive Promoters for Applications in Pathway Engineering in Saccharomyces cerevisiae.pdf

Media:Tum12_Engineering_of_Promoter_Replacement_Cassettes_for_Fine-Tuning_of_Gene_Expression_in_Saccharomyces_cerevisiae.pdf‎

@@ Line 8: / Line 8: @@
 [[File:Georg_einzel_TUM12.jpg|200px|thumb||Responsible: Georg Schützinger]]
-Motiviational Text why we need constitutive promoters and why we explicitly chose those three.
+For the expression of of proteins like limonene, thaumatin and others promoters are essential.
 ==Background and principles==

Team:TU Munich/Project/Constitutive Promoter

From 2012.igem.org