Back
Inducible, tightly regulated and growth condition-independent transcription factor in Saccharomyces cerevisiae.
Journal article

Inducible, tightly regulated and growth condition-independent transcription factor in Saccharomyces cerevisiae.

  • Ottoz DS ETH Zurich and Swiss Institute of Bioinformatics, D-BSSE, Mattenstrasse 26, 4058 Basel, Switzerland Life Science Zurich PhD Program on Molecular and Translational Biomedicine, Zurich, Switzerland Competence Center for Systems Physiology and Metabolic Diseases, ETH Zurich, 8093 Zurich, Switzerland.
  • Rudolf F ETH Zurich and Swiss Institute of Bioinformatics, D-BSSE, Mattenstrasse 26, 4058 Basel, Switzerland joerg.stelling@bsse.ethz.ch.
  • Stelling J ETH Zurich and Swiss Institute of Bioinformatics, D-BSSE, Mattenstrasse 26, 4058 Basel, Switzerland Competence Center for Systems Physiology and Metabolic Diseases, ETH Zurich, 8093 Zurich, Switzerland joerg.stelling@bsse.ethz.ch.
  • 2014-07-19
Published in:
  • Nucleic acids research. - 2014
Bacterial Proteins
Gene Expression Regulation
Genetic Engineering
Humans
Promoter Regions, Genetic
Protein Structure, Tertiary
Receptors, Estrogen
Recombinant Fusion Proteins
Saccharomyces cerevisiae
Serine Endopeptidases
Transcription, Genetic
English The precise control of gene expression is essential in basic biological research as well as in biotechnological applications. Most regulated systems available in yeast enable only the overexpression of the target gene, excluding the possibility of intermediate or weak expression. Moreover, these systems are frequently toxic or depend on growth conditions. We constructed a heterologous transcription factor that overcomes these limitations. Our system is a fusion of the bacterial LexA DNA-binding protein, the human estrogen receptor (ER) and an activation domain (AD). The activity of this chimera, called LexA-ER-AD, is tightly regulated by the hormone β-estradiol. The selection of the AD proved to be crucial to avoid toxic effects and to define the range of activity that can be precisely tuned with β-estradiol. As our system is based on a heterologous DNA-binding domain, induction in different metabolic contexts is possible. Additionally, by controlling the number of LexA-binding sites in the target promoter, one can scale the expression levels up or down. Overall, our LexA-ER-AD system is a valuable tool to precisely control gene expression in different experimental contexts without toxic side effects.
Language
  • English
Open access status
gold
Identifiers
Persistent URL
https://sonar.ch/global/documents/143401
Statistics
Document views: 47 File downloads: