| Literature DB >> 28423198 |
Makoto J Ohira1, David G Hendrickson2, R Scott McIsaac2, Nicholas Rhind1.
Abstract
The fission yeast Schizosaccharomyces pombe lacks a diverse toolkit of inducible promoters for experimental manipulation. Available inducible promoters suffer from slow induction kinetics, limited control of expression levels and/or a requirement for defined growth medium. In particular, no S. pombe inducible promoter systems exhibit a linear dose-response, which would allow expression to be tuned to specific levels. We have adapted a fast, orthogonal promoter system with a large dynamic range and a linear dose response, based on β-estradiol-regulated function of the human oestrogen receptor, for use in S. pombe. We show that this promoter system, termed Z3 EV, turns on quickly, can reach a maximal induction of 20-fold, and exhibits a linear dose response over its entire induction range, with few off-target effects. We demonstrate the utility of this system by regulating the mitotic inhibitor Wee1 to create a strain in which cell size is regulated by β-estradiol concentration. This promoter system will be of great utility for experimentally regulating gene expression in fission yeast.Entities:
Keywords: Schizosaccharomyces; ZEV; estradiol; fission yeast; inducible promoter; pombe
Mesh:
Substances:
Year: 2017 PMID: 28423198 PMCID: PMC5542879 DOI: 10.1002/yea.3235
Source DB: PubMed Journal: Yeast ISSN: 0749-503X Impact factor: 3.239