[Construction and characterization of an hcp mutant of swine Bordetella bronchiseptica].

微生物学报 Pub Date : 2017-02-04
Yuhui Chang, Hongfeng Zhang, Zhong Peng, Hui Li, Huanchun Chen, Bin Wu
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引用次数: 0

Abstract

Objective: We constructed Bordetella bronchiseptica QH0814 hcp mutant to characterize its pathogenicity.[Methods] Through the homologous recombination mediated by a suicide plasmid pRE112, we acquired the mutant QH0814Δhcp successfully. Then, we evaluated the growth condition, the ability of adhesion and invasion, the median lethal dose (LD50) and the infection capacity.

Methods: Through the homologous recombination mediated by a suicide plasmid pRE112, we acquired the mutant QH0814Δhcp successfully. Then, we evaluated the growth condition, the ability of adhesion and invasion, the median lethal dose (LD50) and the infection capacity.

Results: There was no significant variation of the growth rate between the mutant and the parental strain. Compared with the parental strain, the adherence ability of the mutant did not change remarkably. However, the invasion ability decreased significantly. Mice lethal test showed that the LD50 of the mutant was higher than that of the parental strain. Correspondingly, the bacterial colonization of the mutant in mice blood, lung and liver was much less than that of the parental strain.

Conclusion: The knocking-out of the hcp gene had no influence on bacterial growth, but it could attenuate significantly the invasion and colonization of the bacterium. Therefore, the gene may play a role in the pathogenesis of Bordetella bronchiseptica.

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[猪支气管杆菌hcp突变体的构建和鉴定]。
目的:构建支气管脓毒杆菌QH0814 hcp突变体,研究其致病性。[方法]通过自杀质粒pRE112介导的同源重组,成功获得突变体QH0814Δhcp。然后对其生长条件、粘附和侵袭能力、中位致死剂量(LD50)和感染能力进行评价。方法:通过自杀质粒pRE112介导的同源重组,成功获得突变体QH0814Δhcp。然后对其生长条件、粘附和侵袭能力、中位致死剂量(LD50)和感染能力进行评价。结果:突变体与亲本间生长速率无显著差异。与亲本相比,突变体的粘附能力没有明显变化。然而,入侵能力明显下降。小鼠致死试验表明,该突变株的LD50高于亲本株。相应地,突变菌株在小鼠血液、肺和肝脏中的细菌定植量远低于亲本菌株。结论:敲除hcp基因对细菌生长无影响,但能明显减弱细菌的侵袭和定植。因此,该基因可能在支气管脓毒杆菌的发病机制中起作用。
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期刊介绍: Acta Microbiologica Sinica(AMS) is a peer-reviewed monthly (one volume per year)international journal,founded in 1953.It covers a wide range of topics in the areas of general and applied microbiology.The journal publishes original papers,reviews in microbiological science,and short communications describing unusual observations. Acta Microbiologica Sinica has been indexed in Index Copernicus (IC),Chemical Abstract (CA),Excerpt Medica Database (EMBASE),AJ of Viniti (Russia),Biological Abstracts (BA),Chinese Science Citation Database (CSCD),China National Knowledge Infrastructure(CNKI),Institute of Scientific and Technical Information of China(ISTIC),Chinese Journal Citation Report(CJCR),Chinese Biological Abstracts,Chinese Pharmaceutical Abstracts,Chinese Medical Abstracts and Chinese Science Abstracts.
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