Characterization of different promoters for designing a new expression vector in Saccharomyces cerevisiae

Yeast
Siavash PartowJérôme Maury

Abstract

The widely used pESC vector series (Stratagene, La Jolla, CA, USA) with the bidirectional GAL1/GAL10 promoter provides the possibility of simultaneously expressing two different genes from a single vector in Saccharomyces cerevisiae. This system can be induced by galactose and is repressed by glucose. Since S. cerevisiae prefers glucose as a carbon source, and since its growth rate is higher in glucose than in galactose-containing media, we compared and evaluated seven different promoters expressed during growth on glucose (pTEF1, pADH1, pTPI1, pHXT7, pTDH3, pPGK1 and pPYK1) with two strong galactose-induced promoters (pGAL1 and pGAL10), using lacZ as a reporter gene and measuring LacZ activity in batch and continuous cultivation. TEF1 and PGK1 promoters showed the most constant activity pattern at different glucose concentrations. Based on these results, we designed and constructed two new expression vectors which contain the two constitutive promoters, TEF1 and PGK1, in opposite orientation to each other. These new vectors retain all the features from the pESC-URA plasmid except that gene expression is mediated by constitutive promoters.

References

Sep 1, 1990·Molecular and Cellular Biology·J S Flick, M Johnston
Oct 25, 1981·Journal of Molecular Biology·T P St John, R W Davis
Nov 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·A Laughon, R F Gesteland
Mar 25, 1983·Nucleic Acids Research·R DerynckD V Goeddel
Jan 24, 1998·Genome Research·N ErdenizR Rothstein
May 21, 1999·The Journal of Biological Chemistry·J A DiderichA L Kruckeberg
Jun 21, 2002·Methods in Enzymology·R Daniel Gietz, Robin A Woods
Mar 22, 2007·Microbial Cell Factories·María J QuinteroSebastián Chávez
Aug 21, 2007·Biotechnology and Bioengineering·Mohammad A AsadollahiJens Nielsen

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Citations

May 19, 2011·Nucleic Acids Research·Nathan C CrookHal S Alper
Aug 21, 2012·Applied and Environmental Microbiology·Kuk-Ki Hong, Jens Nielsen
Feb 19, 2013·Annual Review of Chemical and Biomolecular Engineering·Yen-Hsiang WangChristina D Smolke
May 17, 2011·Microbial Cell Factories·William A Rodríguez-LimasLaura A Palomares
Jan 10, 2014·Microbial Cell Factories·Katrin WeinhandlAndrea Camattari
Sep 10, 2013·Applied and Environmental Microbiology·Xuelei TianWei Xiao
Feb 9, 2012·FEBS Letters·Benjamin A BlountTom Ellis
Oct 4, 2011·Biotechnology Advances·Eda Celik, Pınar Calık
Jan 7, 2016·Applied Biochemistry and Biotechnology·Yan LiPingkai Ouyang
Nov 3, 2015·Biochemical and Biophysical Research Communications·Tomonao Inobe, Reiko Genmei
Mar 7, 2013·Molecular Microbiology·Kentaro Furukawa, Stefan Hohmann
Mar 5, 2013·Biotechnology and Bioengineering·Kanokarn KocharinJens Nielsen
Feb 18, 2011·Biotechnology Journal·Anastasia KrivoruchkoJens Nielsen
May 20, 2015·Metabolic Engineering·Isis J Trenchard, Christina D Smolke
Dec 20, 2011·Biotechnology and Bioengineering·Zihe LiuJens Nielsen
Mar 27, 2012·Chembiochem : a European Journal of Chemical Biology·Kan'ichiro IshiuchiKenji Watanabe
Mar 5, 2014·Plant, Cell & Environment·Claudia E VickersHarro Bouwmeester
Mar 29, 2014·Biotechnology Journal·Irina Borodina, Jens Nielsen
Sep 18, 2015·Journal of Industrial Microbiology & Biotechnology·Vratislav StovicekIrina Borodina
Nov 26, 2015·Scientific Reports·Nan ShaoMohan K Balasubramanian
Nov 5, 2013·Trends in Biotechnology·Oliver SpadiutChristoph Herwig
Nov 5, 2014·Biotechnology and Bioengineering·Ju Young LeeKwang Myung Cho
Jun 14, 2014·Biochemical and Biophysical Research Communications·Sonoko KawamuraJiro Toshima
Dec 7, 2013·Fungal Genetics and Biology : FG & B·Tiantian WangXiaomin Zhao

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