布鲁氏菌ery操纵子突变株生物学特性分析  

Biological Characterization of an ery Operon Promoter Mutant in Brucella

作　　者：桂丹[1] 张辉[1] 孟仁[1] 孟茹[1] 张豫[1] 孙志华[1] 蒋攀文[1] 李天森[1] 陈创夫[1] 

机构地区：[1]石河子大学动物科技学院,新疆石河子832000

出　　处：《中国生物化学与分子生物学报》2013年第1期76-83,共8页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家重点基础研究发展规划(973项目;No.2010CB30203);国家自然科学基金(No.31001046/30960288);高层次人才科研启动资金(No.RCZX200914)~~

摘　　要：布鲁氏菌ery操纵子参与赤藓醇代谢.赤藓醇能够促进布鲁氏菌的生长.为进一步研究布鲁氏菌引发宿主流产的分子机制,采用基因重组技术构建布鲁氏菌ery操纵子启动子缺失株(△ery),通过体内外实验探讨布鲁氏菌ery操纵子的生物学功能.研究结果显示,获得了布鲁氏菌ery操纵子缺失株;布鲁氏菌ery操纵子缺失株侵染胚胎滋养层细胞脱落较亲本株明显下降;巨噬细胞CFU计数缺失株作用组和亲本株作用组差异显著(P<0.05).试管凝集和虎红平板实验结果显示均出现凝集现象;检测血清中细胞因子IL-10和TNF-α的表达水平,△ery诱导机体产生的IL-10和TNF-α明显低于亲本株(P<0.05).小鼠脾脏细菌CFU计数结果显示,△ery较亲本株毒力明显下降.本研究表明,布鲁氏菌ery操纵子启动子缺失株毒力较亲本株明显下降,为进一步揭示布鲁氏菌引起流产的致病机制提供了一定的理论依据.The ery operon is important for the metabolism of erythritol in Brucella, and erythritol was known to promote the growth of Brucella. To study the molecular mechanisms of Brucella-induced host abortion, an ery operon promoter deletion mutant strain （△ery） was constructed by DNA recombination to explore the biological function of Brucella ery operon in vivo and in vitro. The results showed that the exfoliative human trophoblast cells （HTP-8） infected with the △ery strain was significantly decreased as compared those infected with the wild type in clone forming units （CFU） （P〈0.05）. In tube agglutination and rose bengal plate agglutination tests, the results showed that the degree of agglutination, as well as the levels of IL-10 and TNF-α were significantly lower in △ery groups （P〈0.05） than in the controls. The mouse spleen CFU results showed that the virulence of △ery was significantly decreased. This study provided useful evidence indicating the ery operon could be important for the pathogenesis of Brucella in abortion.

关 键 词：布鲁氏菌 ery操纵子 缺失株 毒力 

分 类 号：S852[农业科学—基础兽医学]