title: An IPTG-inducible derivative of the fission yeast nmt promoter.
abstract: We here describe an IPTG-inducible system that reveals that the lac repressor alone can function as a potent transmodulator to regulate gene expression in the fission yeast, Schizosaccharomyces pombe. This expression system is a derivative of the Sz. pombe nmt promoter, which normally is strongly repressed by thiamine. With appropriate positioning of a lac operator site (lacO) downstream of the TATA-box, we show that gene expression from a chimeric nmt::lacO promoter can be regulated by the lac repressor up to two orders of magnitude in response to IPTG. The chimeric nmt::lacO promoter is rapidly induced and when GFP is used as a reporter; almost full induction is achieved 40 min after the addition of IPTG. Like the wild-type nmt promoter, the chimeric nmt::lacO is repressed by thiamine. This allows expression in a short and defined window, e.g. the S-phase of a synchronized cell population, by first adding IPTG to turn on expression, followed by addition of thiamine to switch off expression.
authors: Department of Biology, University of Copenhagen, DK-2220, Copenhagen N, Denmark.|Chemometec, DK-3450, Allerod, Denmark.
authors: Søren
authors: S
authors: Kjaerulff
authors: Department of Biology, University of Copenhagen, DK-2220, Copenhagen N, Denmark.
authors: Olaf
authors: O
authors: Nielsen
mesh_terms: D015966:Gene Expression Regulation, Fungal; D007544:Isopropyl Thiogalactoside / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D012568:Schizosaccharomyces / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics* / Q000378:metabolism; D013831:Thiamine / Q000378:metabolism
chemical_list: D012097:Repressor Proteins; D029702:Schizosaccharomyces pombe Proteins; D007544:Isopropyl Thiogalactoside; D013831:Thiamine
keywords: IPTG; Sz. pombe; expression system; lacI; lacO; nmt