This study describes a PCR-based approach for the production of a rationally attenuated mutant of Degenerate primers were used to amplify a fragment encoding 91.45% of the gene of MP6 which was cloned into pUC18. The remainder of the gene was isolated by inverse PCR. The gene was sequenced and a restriction map was generated. The gene had 75.9% identity with the gene of The cloned gene was inactivated and reintroduced into strain GB using the suicide vector pGP704. A stable -defective mutant, GBΔ was isolated and its virulence was examined The mutant was attenuated in guinea-pigs and capable of inducing a protective immune response against challenge with the virulent strain GB. Unusually for an -defective mutant, the mutant was virulent in mice, with a median dose which induced morbidity or death similar to that of the wild-type, although time to death was significantly prolonged.

Keyword(s): aroA and Yersinia pestis

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