Wild-type strains of Dictyostelium discoideum can be transformed using a novel selection cassette driven by the promoter of the ribosomal V18 gene.
Almost all methods for transformation of the social ameba Dictyostelium discoideum rely on axenic growth, that is, growth in a synthetic medium, for at least part of the procedure. Axenic growth requires several mutations. Here we describe a procedure that can be used to transform wild-type strains which are able to grow only on the natural food source, bacteria. The method relies on a new selection cassette driven by the V18 promoter, a promoter that we show is substantially more active during growth on bacteria than the actin-6 promoter, which is widely used for axenic transformation. The procedure gives transformation frequencies of about 10(-5) with both strains Ax2 (capable of axenic growth) and NC4 (capable of growth only on bacteria). Using this vector, we have obtained NC4 strains carrying several beta-galactosidase reporter cassettes. Our vector can also be used in axenic transformations.
Wetterauer
B
B
Zoologisches Institut, Ludwig-Maximilians-Universität, Luisenstrasse 14, Munich, 80333, Germany.
Morandini
P
P
Hribar
I
I
Murgia-Morandini
I
I
Hamker
U
U
Singleton
C
C
Macwilliams
H K
HK
D000199:Actins; D000818:Animals; D004254:DNA Nucleotidyltransferases; D004023:Dictyostelium / Q000235:genetics* / Q000254:growth & development; D007711:Klebsiella pneumoniae; D011401:Promoter Regions, Genetic; D012269:Ribosomal Proteins / Q000235:genetics*; D014170:Transformation, Genetic; D019895:Transposases
D000199:Actins; D012269:Ribosomal Proteins; D004254:DNA Nucleotidyltransferases; D019895:Transposases