Use of a transactive regulatory mutant of Dictyostelium discoideum in a eucaryotic expression system.
The discoidin proteins of Dictyostelium discoideum are highly expressed during development. The Disc I gamma promoter allows the regulation of heterologous protein expression by experimental conditions. 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 discoidins also overexpresses the reporter genes beta-galactosidase, 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 overexpression of genes in Dictyostelium, both for functional studies in vivo and for the production of heterologous proteins for purification.
Wetterauer
B W
BW
Zoologisches Institut, Ludwig-Maximilians Universität München, Germany.
Salger
K
K
MacWilliams
H K
HK
D000818:Animals; D003001:Cloning, Molecular / Q000379:methods; D004023:Dictyostelium / Q000235:genetics*; D000070821:Discoidins; D005656:Fungal Proteins / Q000235:genetics*; D005786:Gene Expression Regulation; D005800:Genes, Fungal; D037102:Lectins; D009154:Mutation; D011401:Promoter Regions, Genetic; D015800:Protozoan Proteins; D012333:RNA, Messenger / Q000235:genetics
D000070821:Discoidins; D005656:Fungal Proteins; D037102:Lectins; D015800:Protozoan Proteins; D012333:RNA, Messenger