A System for Temperature-Controlled Expression of a Foreign Gene with Dual Mode in Saccharomyces cerevisiae

概要

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A temperature-controlled expression system for a foreign gene in Saccharomyces cerevisiae was constructed. In this system, a MATa hmlaα2-102 HMRa sir3-8^<ts> double mutant was used as host, and a DNA fragment bearing the promoter and pre- or pro-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℃ to 35℃) 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℃), 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℃ to 30℃. 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.
社団法人日本生物工学会の論文
1990-06-25