Construction of Temperature–Regulated Yeast Promoters Using the Matα2 Repression System

Abstract

Temperature–regulated hybrid yeast promoters were constructed in which synthetic MATα2 operator sequences were inserted into the strong promoters for the constitutive TPI1 (triose phosphate isomerase) gene and the glucose–repressible ADH2 (alcohol dehydrogenase) gene. These variant promoters were joined to the E. coli lacZ gene in yeast vectors and transformed into mating–type a yeast strains bearing a temperature–sensitive sir3–8 mutation. At the permissive temperature (25°C), the sir3–8 product represses the silent mating–type loci HMRa and HMLα, so that the MATα2 represser protein is not synthesized by HMLα and β–galactosidase is produced at high levels. At the restrictive temperature (35°C), MATα2 repressor is made and represses transcription of lacZ. At temperatures between 25°C and 35°C, intermediate levels of β–galactosidase are made. Thus, the level of heterologous product can be altered by adjusting the temperature.

Cite this article

Sledziewski, A., Bell, A., Kelsay, K. et al. Construction of Temperature–Regulated Yeast Promoters Using the Matα2 Repression System. Nat Biotechnol 6, 411–416 (1988). https://doi.org/10.1038/nbt0488-411

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