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An estradiol‐inducible promoter enables fast,graduated control of gene expression in fission yeast
Authors:Makoto J Ohira  David G Hendrickson  R Scott McIsaac  Nicholas Rhind
Affiliation:1. Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA, USA;2. Calico Life Sciences, South San Francisco, CA, USA
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 Z3EV, 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. Copyright © 2017 John Wiley & Sons, Ltd.
Keywords:Schizosaccharomyces  pombe  fission yeast  ZEV  inducible promoter  estradiol
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