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Evaluation of the Saccharomyces cerevisiae ADH2 promoter for protein synthesis
Author(s) -
Michael Lee K.,
DaSilva Nancy A.
Publication year - 2005
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.1221
Subject(s) - derepression , biology , saccharomyces cerevisiae , promoter , inducer , lac operon , yeast , transcription (linguistics) , plasmid , gene , microbiology and biotechnology , recombinant dna , biochemistry , gene expression , psychological repression , linguistics , philosophy
The Saccharomyces cerevisiae ADH2 promoter (P ADH2 ) is repressed several hundred‐fold in the presence of glucose; transcription is initiated once the glucose in the medium is exhausted. The promoter can thus be utilized for effective regulation of recombinant gene expression in S. cerevisiae without the addition of an inducer. To evaluate this promoter in the absence of plasmid copy number and stability variations, the P ADH2 − lacZ cassette was integrated into the yeast chromosomes. The effects of medium composition, glucose concentration and cultivation time on promoter derepression and expression level were investigated. Maximum protein activity was obtained after 48 h of growth in complex YPD medium containing 1% glucose. The widely used S. cerevisiae GAL1 and CUP1 promoters both require the addition of an inducer [galactose and copper(II) ion, respectively] before regulated genes will be expressed. The strengths of these three different promoters were compared for cells containing one copy of an integrated lacZ gene under their control. The ADH2 promoter was superior for all induction strategies investigated. Copyright © 2005 John Wiley & Sons, Ltd.