Characterization of different promoters for designing a new expression vector in Saccharomyces cerevisiae
Artikel i vetenskaplig tidskrift, 2010

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.

phosphoglycerate kinase

heterologous gene-expression

yeast

fluorescent protein

glyceraldehyde-3-phosphate dehydrogenase

bidirectional promoter

promoter activity

individual hexose transporters

gal1 promoter

green

expression vector

pgk-gene

yeast

glucose repression

hxt genes

Författare

Siavash Partow

Kemi- och bioteknik, Livsvetenskaper, Systembiologi

Verena Siewers

Kemi- och bioteknik, Livsvetenskaper, Systembiologi

S. Bjorn

Fluxome Sciences A/S

Jens B Nielsen

Kemi- och bioteknik, Livsvetenskaper, Systembiologi

J. Maury

Fluxome Sciences A/S

Yeast

0749-503X (ISSN) 1097-0061 (eISSN)

Vol. 27 955-964

Ämneskategorier

Industriell bioteknik

Mikrobiologi

Styrkeområden

Livsvetenskaper och teknik

DOI

10.1002/yea.1806