Molecular analysis of the aroA gene of Pasteurella multocida and vaccine potential of a constructed aroA mutant.
The aroA gene from Pasteurella multocida was cloned by complementation of the Escherichia coli aroA mutant AB2829 with a DNA library constructed in pUC18. The nucleotide sequence of the P. multocida aroA gene indicated an open reading frame encoding a protein of 441 amino acids, which showed a high degree of homology with the amino acid sequences of various other bacterial AroA proteins. The cloned P. multocida aroA gene was inactivated by insertion of a kanamycin-resistance gene and reintroduced by allelic exchange into the chromosome of P. multocida using the suicide vector pJM703.1. The P. multocida aroA mutant was highly attenuated in a mouse model. Mice immunized intraperitoneally with two doses of live P. multocida aroA mutant were completely protected against a lethal parental strain challenge.
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Department of Microbiology, Monash University, Clayton, Victoria, Australia.
Strugnell
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Adler
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D000595:Amino Acid Sequence; D000818:Animals; D001428:Bacterial Vaccines / Q000235:genetics*; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D005260:Female; D005798:Genes, Bacterial; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D016979:Pasteurella multocida / Q000235:genetics* / Q000472:pathogenicity; D014613:Vaccines, Attenuated / Q000235:genetics; D014614:Vaccines, Synthetic / Q000235:genetics*
D001428:Bacterial Vaccines; D004269:DNA, Bacterial; D014613:Vaccines, Attenuated; D014614:Vaccines, Synthetic