Construction and vaccine potential of an aroA mutant of Pasteurella haemolytica.
The aroA gene, encoding 5-enolpyruvylshikimate 3-phosphate synthase, from Pasteurella haemolytica biotype A, serotype 1 was cloned by complementation of the aroA mutation in Escherichia coli strain AB2829 after electroporation with a DNA library constructed in pUC18. The cloned P. haemolytica aroA gene was inactivated by insertion of a kanamycin resistance gene and reintroduced by allelic exchange into the chromosome of the parental P. haemolytica using PbluescriptII SK+. The P. haemolytica aroA mutant was highly attenuated in a mouse septicaemic model. Mice immunized intraperitoneally with two doses of live P. haemolytica aroA mutant were protected against a lethal parental strain challenge.
Department of Microbiology, Monash University, Clayton, Melbourne, Vic., Australia.
P
P
Homchampa
R A
RA
Strugnell
B
B
Adler
D000818:Animals; D016470:Bacteremia / Q000382:microbiology / Q000517:prevention & control; D015373:Bacterial Typing Techniques / Q000662:veterinary; D001428:Bacterial Vaccines / Q000235:genetics*; D015139:Blotting, Southern / Q000662:veterinary; D003001:Cloning, Molecular; D004195:Disease Models, Animal; D018274:Electroporation / Q000662:veterinary; D004926:Escherichia coli; D005260:Female; D007124:Immunoenzyme Techniques / Q000662:veterinary; D016978:Mannheimia haemolytica / Q000235:genetics* / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009154:Mutation / Q000276:immunology*; D010326:Pasteurella Infections / Q000517:prevention & control / Q000662:veterinary*; D012766:Pasteurellosis, Pneumonic / Q000517:prevention & control; D017422:Sequence Analysis, DNA / Q000662:veterinary; D012703:Serotyping / Q000662:veterinary
D001428:Bacterial Vaccines