Generation of attenuated Mycobacterium bovis strains by signature-tagged mutagenesis for discovery of novel vaccine candidates

Desmond Collins, Bronwyn Skou, Stefan John White, Shalome Bassett, Lauren Collins, Raewyn For, Kathryn Hurr, Grant Hotter, Geoffrey de Lisle

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23 Citations (Scopus)

Abstract

Mycobacterium bovis, a member of the Mycobacterium tuberculosis complex, has a particularly wide host range and causes tuberculosis in most mammals, including humans. A signature tag mutagenesis approach, which employed illegitimate recombination and infection of guinea pigs, was applied to M. bovis to discover genes important for virulence and to find potential vaccine candidates. Fifteen attenuated mutants were identified, four of which produced no lesions when inoculated separately into guinea pigs. One of these four mutants had nine deleted genes including mmpL4 and sigK and, in guinea pigs with aerosol challenge, provided protection against tuberculosis at least equal to that of M. bovis BCG. Seven mutants had mutations near the esxA (esat-6) locus, and immunoblot analysis of these confirmed the essential role of other genes at this locus in the secretion of EsxA (ESAT-6) and EsxB (CFP10). Mutations in the eight other attenuated mutants were widely spread through the chromosome and included pks1, which is naturally inactivated in clinical strains of M. tuberculosis. Many genes identified were different from those found by signature tag mutagenesis of M. tuberculosis by use of a mouse infection model and illustrate how the use of different approaches enables identification of a wider range of attenuating mutants.
Original languageEnglish
Pages (from-to)2379 - 2386
Number of pages8
JournalInfection and Immunity
Volume73
Issue number4
DOIs
Publication statusPublished - 2005
Externally publishedYes

Cite this

Collins, D., Skou, B., White, S. J., Bassett, S., Collins, L., For, R., Hurr, K., Hotter, G., & de Lisle, G. (2005). Generation of attenuated Mycobacterium bovis strains by signature-tagged mutagenesis for discovery of novel vaccine candidates. Infection and Immunity, 73(4), 2379 - 2386. https://doi.org/10.1128/IAI.73.4.2379-2386.2005