A system for temperature-controlled expression of a foreign gene with dual mode in Saccharomyces cerevisiae

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Abstract A temperature-controlled expression system for a foreign gene in Saccharomyces cerevisiae was constructed. In this system, a MAT a hmlα2-102 HMR a sir3–8 ts double mutant was used as host, and a DNA fragment bearing the promoter and pre- or pre-pro regions of the MFα1 gene encoding the α-factor of S. cerevisiae was used as a promoter for expression of a foreign gene cloned on a vector. When the host cells were incubated at a restrictive temperature for the sir3–8 ts mutation (30°C to 35°C) they showed the α mating type and a PHO5 DNA fragment of S. cerevisiae , encoding repressible acid phosphatase, connected downstream of the MFα1 promoter was expressed. But when they were incubated at permissive lower temperature (25°C), at which they have the a mating type, the PHO5 DNA was not expressed. Acid phosphatase activity was increased 30-fold by shifting the incubation temperature from 25°C to 30°C. In this system it may also be possible to express a foreign gene at lower temperature but shut off its expression at higher temperature by connecting the gene to a promoter DNA of an a-specific gene.

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