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【目的】构建猪支气管败血波氏杆菌(Bordetella bronchiseptica,Bb)Ⅵ型分泌系统(T6SS)溶血素共调节蛋白hcp基因缺失株,并对其基本生物学特性进行初步的研究。【方法】使用自杀性质粒介导同源重组的方法敲除猪支气管败血波氏杆菌QH0814菌株hcp基因,并比较hcp基因缺失前后,菌体对细胞的黏附入侵、小鼠毒力及组织载菌量上的差异。【结果】成功构建支气管败血波氏杆菌hcp基因缺失株QH0814Δhcp,连续传50代且遗传稳定;缺失株与亲本株生长无明显差异;缺失株的黏附能力与亲本株差异不显著,但入侵能力显著降低(P<0.05);与亲本株相比,半数致死量提高,同时,缺失株对昆明鼠的感染能力也显著降低(P<0.05)。【结论】hcp基因的缺失对支气管败血波氏杆菌增殖无影响,但缺失后其入侵能力和定殖能力显著降低,由此推测hcp基因与支气管败血波氏杆菌的入侵和定殖相关。
【Objective】 To construct the hcp gene deletion mutant of pig T6SS pig bordetella bronchiseptica (Bb), and to study its basic biological characteristics. 【Method】 Hcp gene was knocked out by using homologous recombination mediated by suicide plasmid. The hcp gene of porcine Bordetella bronchiseptica QH0814 was knocked out and compared before and after deletion of hcp gene, Differences in the amount of bacteria. 【Result】 The results showed that QH0814Δhcp gene was successfully constructed in Hcp gene of Bordetella bronchiseptica and its inheritance was stable for 50 passages. There was no significant difference in growth between the deletion strain and the parental strain, but no significant difference in the adhesion ability between the deletion strain and the parental strain. However, (P <0.05). Compared with the parental strain, the median lethal dose increased and the ability of the mutant strain to infect Kunming mice significantly decreased (P <0.05). 【Conclusion】 The deletion of hcp gene has no effect on the proliferation of Bordetella bronchiseptica, but its abilities of invasion and colonization are significantly reduced after deletion, suggesting that hcp gene is associated with invasion and colonization of B. bronchiseptica.