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Nucleic Acids Research, 1993, Vol. 21, No. 6 1397-1401
© 1993

METHODS

Use of a transactive regulatory mutant of Dictyostelium discoideum in a eucaryotic expression system

Birgit W. Wetterauer, Klaus Salger and Harry K. MacWilliams

Zoologisches Instrtut der Ludwig-Maximilians Universität München Luisenstraße 14, 8000 München 2, Germany

Received January 4, 1993. Revised February 15, 1993. Accepted February 15, 1993.

The discoldin proteins of Dictyostellum dlscoideum are highly expressed during development. The Disc I{gamma} promoter allows the regulation of heterologous protein expression by experimental conditions (1). We report conditions under which the promoter activity is efficiently repressed during growth In the wildtype strain AX2. In addition we show that a mutant which overexpresses the discoidlns also overexpresses the reporter genes ß-galactosldase, luciferase and CAT 10-to 100-fold when these are placed under the control of a Disc I{gamma} promoter. This system may be generally useful for the overexpresslon of genes In Dictyostellum, both for functional studies In vivo and for the production of heterologous proteins for purification.


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