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Methods Mol Biol. 2006;346:187-99.

Generation of multiple knockout mutants using the Cre-loxP system.

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Laboratory of Cellular and Developmental Biology, NIDDK, National Institutes of Health, Bethesda, MD, USA.


Dictyostelium discoideum is an exceptionally powerful system by which to study various aspects of modern cell and developmental biology. The completion of the genome sequencing project together with a high-efficiency of targeted gene disruption have enabled researchers to characterize many specific gene functions. However, as a result of many gene products with overlapping functions, there is great need to produce mutants carrying mutations in multiple genes. We have, therefore, developed a robust system and describe a protocol for the generation of multiple gene mutations in Dictyostelium by recycling the Blasticidin S selectable marker after transient expression of Cre recombinase.

[Indexed for MEDLINE]

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