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FEMS Microbiol Lett. 1996 Nov 1;144(2-3):161-6.

Positive selection of allelic exchange mutants in Mycobacterium bovis BCG.

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1
Unité de Génétique Mycobactèrienne, Institut Pasteur, Paris, France.

Abstract

sacB expression is lethal to mycobacteria in the presence of sucrose. It can therefore serve as 1 counter-selectable marker for positive selection of gene replacement events as demonstrated in the fast-growing Mycobacterium smegmatis. With this methodology, a sucrose counter-selectable vector was used to deliver, into the Mycobacterium bovis BCG genome, an inactivated copy (ureC::Km) of the ureC gene encoding the mycobacterial urease. A two-step selection procedure on 2% sucrose allowed the positive selection of gene exchange mutants. This technique should thus be extremely useful for the genetic analysis of pathogenic mycobacteria.

[Indexed for MEDLINE]

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