Plasmid. 1996 Nov;36(3):169-81.

Wild-type strains of Dictyostelium discoideum can be transformed using a novel selection cassette driven by the promoter of the ribosomal V18 gene.

Wetterauer B, Morandini P, Hribar I, Murgia-Morandini I, Hamker U, Singleton C, Macwilliams HK.

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Zoologisches Institut, Ludwig-Maximilians-Universität, Luisenstrasse 14, Munich, 80333, Germany.

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.

PMID:: 9007012; [PubMed - indexed for MEDLINE]

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