title: Wild-type strains of Dictyostelium discoideum can be transformed using a novel selection cassette driven by the promoter of the ribosomal V18 gene.
abstract: 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.
authors.affiliation: Zoologisches Institut, Ludwig-Maximilians-Universität, Luisenstrasse 14, Munich, 80333, Germany.
authors.forename: B
authors.initials: B
authors.lastname: Wetterauer
authors.forename: P
authors.initials: P
authors.lastname: Morandini
authors.forename: I
authors.initials: I
authors.lastname: Hribar
authors.forename: I
authors.initials: I
authors.lastname: Murgia-Morandini
authors.forename: U
authors.initials: U
authors.lastname: Hamker
authors.forename: C
authors.initials: C
authors.lastname: Singleton
authors.forename: H K
authors.initials: HK
authors.lastname: Macwilliams
mesh_terms: 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
chemical_list: D000199:Actins; D012269:Ribosomal Proteins; D004254:DNA Nucleotidyltransferases; D019895:Transposases