Team:TU Munich/Project/Constitutive Promoter

From 2012.igem.org

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(Background and principles)
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==Background and principles==
==Background and principles==
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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.
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.
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Elke Nevoigt et al. generated a library of TEF1 promotermutants for finetuning of Expression [[Media:Tum12 TEF-Promoter-mutants.pdf]]
Elke Nevoigt et al. generated a library of TEF1 promotermutants for finetuning of Expression [[Media:Tum12 TEF-Promoter-mutants.pdf]]
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==General remarks and issues==
==General remarks and issues==
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Revision as of 21:05, 23 September 2012


Contents

Constitutive Promoters


Responsible: Georg Schützinger

Motiviational Text why we need constitutive promoters and why we explicitly chose those three.

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.

TUM12 Geclonte promoter.jpg


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