f Construction of Bacillus anthracis mutant strains producing a single toxin component
- Authors: Corinne Pezard, Edith Duflot, Michéle Mock*
- *Author for correspondence. Tel. +33 1 45 68 83 12; fax +33 1 45 68 89 54.
- Microbiology, October 1993 139: 2459-2463, doi: 10.1099/00221287-139-10-2459
- Subject: Genetics And Molecular Biology
- Published Online:
SUMMARY: The two protein exotoxins secreted by Bacillus anthvacis are composed of three distinct components: protective antigen (PA), lethal factor (LF), and (o)edema factor (EF). We have developed a genetic strategy that permits us selectively to inactivate each of the genes coding for PA, EF or LF. This strategy involved the deletion of a portion of the structural gene and the insertion of an antibiotic resistance cassette. With this technique, double mutant strains of B. anthracis producing only one toxin component have been constructed. Characterization of the mutant strains indicated that they produced the expected single toxin protein. Using a simple, two-step protocol, we have purified PA, LF and EF to homogeneity from culture supernatants. These three mutant strains are potentially powerful tools for studying the individual effect of each toxin component in vitro and in vivo.
© Society for General Microbiology, 1993 | Published by the Microbiology Society
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