An aroA Mutant of Yersinia Pestis Is Attenuated in Guinea-Pigs, but Virulent in Mice

Microbiology. 1996 Jul;142 ( Pt 7):1847-53. doi: 10.1099/13500872-142-7-1847.

Abstract

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 GB aroA, 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 of death similar to that of the wild-type, although time to death was significantly prolonged.

Publication types

  • Comparative Study

MeSH terms

  • 3-Phosphoshikimate 1-Carboxyvinyltransferase
  • Alkyl and Aryl Transferases*
  • Amino Acid Sequence
  • Animals
  • Bacterial Vaccines / isolation & purification
  • Base Sequence
  • DNA Primers / genetics
  • DNA, Bacterial / genetics
  • Disease Models, Animal
  • Female
  • Guinea Pigs
  • Mice
  • Mice, Inbred BALB C
  • Molecular Sequence Data
  • Mutagenesis
  • Mutation*
  • Plague / etiology
  • Plague / immunology
  • Plague / prevention & control
  • Sequence Homology, Amino Acid
  • Transferases / genetics*
  • Transferases / immunology
  • Vaccines, Attenuated / isolation & purification
  • Virulence / genetics
  • Yersinia pestis / genetics*
  • Yersinia pestis / immunology
  • Yersinia pestis / pathogenicity*

Substances

  • Bacterial Vaccines
  • DNA Primers
  • DNA, Bacterial
  • Vaccines, Attenuated
  • Transferases
  • Alkyl and Aryl Transferases
  • 3-Phosphoshikimate 1-Carboxyvinyltransferase

Associated data

  • GENBANK/L46372