Inactivation of the fliY gene encoding a flagellar motor switch protein attenuates mobility and virulence of Leptospira interrogans strain Lai.
Inactivation of the fliY gene encoding a flagellar motor switch protein attenuates mobility and virulence of Leptospira interrogans strain Lai.
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DOI：
10.1186/1471-2180-9-253
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发表时间：
2009-12-09
期刊：
影响因子：
4.2
通讯作者：
Yan J
中科院分区：
文献类型：
--
作者：
Liao S;Sun A;Ojcius DM;Wu S;Zhao J;Yan J
致病性钩端螺旋体物种引起钩端螺旋体病，一种全球重要的人畜共患疾病。该螺旋体具有活跃的旋转运动，可能有助于病原体在宿主体内的侵入和扩散。FliY是一种鞭毛马达开关蛋白，在其他微生物中控制鞭毛马达方向，但其在钩端螺旋体中的作用，特别是在致病性中的作用仍然未知。构建了问号钩端螺旋体黄疸出血群赖型赖株fliY基因的自杀质粒，并将其插入氨苄青霉素抗性基因（bla）中，经PCR和Western Blot分析证实fliY基因在突变株（fliY-）中失活。失活导致位于同一操纵子中fliY基因下游的fliP和fliQ基因的mRNA缺失。突变株在液体培养基中的旋转运动明显减弱，在半固体培养基中的迁移也明显减弱。与野生型菌株相比，突变株对小鼠巨噬细胞的粘附能力和致炎能力明显降低，对豚鼠的致死率也明显降低。通过本文中使用的方法，fliY的失活显然对下游基因具有极性效应。观察到的表型，包括较低的致病性，可能是一个结果的fliY失活，但也是一个结果的极性效应。
Pathogenic Leptospira species cause leptospirosis, a zoonotic disease of global importance. The spirochete displays active rotative mobility which may contribute to invasion and diffusion of the pathogen in hosts. FliY is a flagellar motor switch protein that controls flagellar motor direction in other microbes, but its role in Leptospira, and paricularly in pathogenicity remains unknown. A suicide plasmid for the fliY gene of Leptospira interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai that was disrupted by inserting the ampicillin resistance gene (bla) was constructed, and the inactivation of fliY gene in a mutant (fliY-) was confirmed by PCR and Western Blot analysis. The inactivation resulted in the mRNA absence of fliP and fliQ genes which are located downstream of the fliY gene in the same operon. The mutant displayed visibly weakened rotative motion in liquid medium and its migration on semisolid medium was also markedly attenuated compared to the wild-type strain. Compared to the wild-type strain, the mutant showed much lower levels of adhesion to murine macrophages and apoptosis-inducing ability, and its lethality to guinea pigs was also significantly decreased. Inactivation of fliY, by the method used in this paper, clearly had polar effects on downstream genes. The phentotypes observed, including lower pathogenicity, could be a consequence of fliY inactivation, but also a consequence of the polar effects.
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影响因子：
5.4
作者：
Liu, Yunying;Zheng, Wei;Yan, Jie
通讯作者：
Yan, Jie
影响因子：
3.1
作者：
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4.4
作者：
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DOI：
10.1073/pnas.89.14.6304
发表时间：
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影响因子：
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通讯作者：
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影响因子：
5.5
作者：
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