RT Journal Article SR Electronic(1) A1 Oyston, Petra C. F. A1 Russell, Paul A1 Williamson, E. Diane A1 Titball, Richard W.YR 1996 T1 An aroA mutant of Yersinia pestis is attenuated in guinea-pigs, but virulent in mice JF Microbiology, VO 142 IS 7 SP 1847 OP 1853 DO https://doi.org/10.1099/13500872-142-7-1847 PB Microbiology Society, SN 1465-2080, AB This study describes a PCR-based approach for the production of a rationally attenuated mutant of Yersinia pestis. Degenerate primers were used to amplify a fragment encoding 91.45% of the aroA gene of Y. pestis 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 Y. pestis aroA gene had 75.9% identity with the aroA gene of Yersinia enterocolitica. The cloned gene was inactivated in vitro and reintroduced into Y. pestis strain GB using the suicide vector pGP704. A stable aro-defective mutant, Y. pestis GBΔaroA, was isolated and its virulence was examined in vivo. The mutant was attenuated in guinea-pigs and capable of inducing a protective immune response against challenge with the virulent Y. pestis strain GB. Unusually for an aro-defective mutant, the Y. pestis aroA 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., UL https://www.microbiologyresearch.org/content/journal/micro/10.1099/13500872-142-7-1847