Chung J Y, Wilkie I, Boyce J D, Townsend K M, Frost A J, Ghoddusi M, Adler B
Bacterial Pathogenesis Research Group, Department of Microbiology, Monash University, Victoria 3800, Australia.
Infect Immun. 2001 Apr;69(4):2487-92. doi: 10.1128/IAI.69.4.2487-2492.2001.
We have constructed a defined acapsular mutant in Pasteurella multocida X-73 (serogroup A:1) by disrupting the hexA gene through the insertion of a tetracycline resistance cassette. The genotype of the hexA::tet(M) strain was confirmed by PCR and Southern hybridization, and the acapsular phenotype of this strain was confirmed by electron microscopy. The hexA::tet(M) strain was attenuated in both mice and chickens. Complementation of the mutant with an intact hexAB fragment restored lethality in mice but not in chickens. In contrast to the results described previously for P. multocida serogroup B (J. D. Boyce and B. Adler, Infect. Immun. 68:3463-3468, 2000), the hexA::tet(M) strain was sensitive to the bactericidal action of chicken serum, whereas the wild-type and complemented strains were both resistant. Following inoculation into chicken muscle, the bacterial count of the hexA::tet(M) strain decreased significantly, while the wild-type and complemented strains both grew rapidly over 4 h. The capsule is thus an essential virulence determinant in the pathogenesis of fowl cholera.
我们通过插入四环素抗性盒破坏多杀性巴氏杆菌X-73(血清群A:1)中的hexA基因,构建了一个明确的无荚膜突变体。通过PCR和Southern杂交确认了hexA::tet(M)菌株的基因型,并通过电子显微镜确认了该菌株的无荚膜表型。hexA::tet(M)菌株在小鼠和鸡中均减毒。用完整的hexAB片段对突变体进行互补,可恢复小鼠的致死性,但不能恢复鸡的致死性。与先前报道的多杀性巴氏杆菌血清群B的结果(J.D. Boyce和B. Adler,《感染与免疫》68:3463-3468,2000)相反,hexA::tet(M)菌株对鸡血清的杀菌作用敏感,而野生型和互补菌株均具有抗性。接种到鸡肌肉中后,hexA::tet(M)菌株的细菌数量显著下降,而野生型和互补菌株在4小时内均快速生长。因此,荚膜是禽霍乱发病机制中一个重要的毒力决定因素。