| Literature DB >> 9007012 |
B Wetterauer1, P Morandini, I Hribar, I Murgia-Morandini, U Hamker, C Singleton, H K Macwilliams.
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.Entities:
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Year: 1996 PMID: 9007012 DOI: 10.1006/plas.1996.0044
Source DB: PubMed Journal: Plasmid ISSN: 0147-619X Impact factor: 3.466