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Properties and engineering of a mutantSTA promoter ofSaccharomyces diastaticus

  • George Bajszár
  • Jon Croonenberghs
  • Irina L. Karnushina
  • Sun Y. Lee
  • James R. Mattoon
Article
  • 28 Downloads

Abstract

A new allelic variant of theSTA2 gene ofS. diastaticus, designated asSTA2 K, was cloned and characterized (1; accompanying paper). An application-oriented analysis of the promoter region ofSTA2 K is described, with an emphasis on its peculiar structural feature: A 1.1-kb natural deletion located 189 nucleotides upstream of the translation start codon.

The strength of theSTA2 K promoter was found comparable to that of known strong constitutive yeast promoters(ADH1, GAPDH). Regulated glucoamylase expression was demonstrated by chimeric promoters, which were constructed by placing theSTA2 K promoter under the control of either thePH05 orCYC1 upstream regulatory sequences. On high-copy-number vectors, induction of the UASpho5-STA2K chimeric promoter by phosphate depletion resulted in a destructive overexpression of the secreted glucoamylase, which completely halted cell growth, and promoted cell decay. In contrast, UAScyc1 was shown to mediate a fine-tuned regulation both by glucose concentration and, indirectly, by starch, the substrate for the glucoamylase to produce glucose.

Index Entries

Yeast glucoamylase promoter, gene expression STA genes Upstream Regulatory Sequences UAS PH05 CYC1 

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Copyright information

© Humana Press Inc 1994

Authors and Affiliations

  • George Bajszár
    • 1
  • Jon Croonenberghs
    • 2
  • Irina L. Karnushina
    • 1
  • Sun Y. Lee
    • 2
  • James R. Mattoon
    • 1
  1. 1.Biotechnology CenterUniversity of Colorado at Colorado SpringsColorado Springs
  2. 2.Coors Brewing Co.Golden

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