RT Journal Article
SR Electronic
T1 An Estradiol-Inducible Promoter Enables Fast, Graduated Control of Gene Expression in Fission Yeast
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 092338
DO 10.1101/092338
A1 Makoto J. Ohira
A1 David G. Hendrickson
A1 R. Scott McIsaac
A1 Nicholas Rhind
YR 2017
UL http://biorxiv.org/content/early/2017/02/17/092338.abstract
AB 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 estrogen 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.