Riemerella anatipestifer M949_1360 Gene Functions on the Lipopolysaccharide Biosynthesis and Bacterial Virulence

文献类型： 外文期刊

第一作者： Yu, Guijing

作者： Yu, Guijing;Wang, Xiaolan;Dou, Yafeng;Wang, Shaohui;Tian, Mingxing;Qi, Jingjing;Li, Tao;Ding, Chan;Yu, Shengqing;Yu, Guijing;Wu, Yantao

作者机构：

期刊名称：PLOS ONE （ 影响因子：3.24； 五年影响因子：3.788 ）

ISSN： 1932-6203

年卷期： 2016 年 11 卷 8 期

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收录情况： SCI

摘要： Riemerella anatipestifer causes septicemic and exudative diseases in poultry, resulting in major economic losses to the duck industry. Lipopolysaccharide (LPS), as an important virulence factor in Gram-negative bacteria, can be recognized by the immune system and plays a crucial role in many interactions between bacteria and animal hosts. In this study, we screened out one LPS defective mutant strain RA Delta 604 from a random transposon mutant library of R. anatipestifer serotype 1 strain CH3, which did not react with the anti-CH3 LPS monoclonal antibody 1C1 in an indirect enzyme-linked immunosorbent assay. Southern blot analysis confirmed that the genome of RA Delta 604 contained a single Tn4351 insert. Then, we found that the M949_1360 gene was inactivated by insertion of the transposon. Using silver staining and western blot analyses, we found that the LPS pattern of RA Delta 604 was defective, as compared with that of the wild-type (WT) strain CH3. The mutant strain RA Delta 604 showed no significant influence on bacterial growth, while bacterial counting and Live/dead BacLight Bacterial Viability staining revealed that bacterial viability was decreased, as compared with the WT strain CH3. In addition, the abilities of the mutant strain RA Delta 604 to adhere and invade Vero cells were significantly decreased. Animal studies revealed that the virulence of the mutant strain RA Delta 604 was decreased by more than 200-fold in a duck infection model, as compared with the WT strain CH3. Furthermore, immunization with live bacteria of the mutant strain RA Delta 604 protected 87.5% ducks from challenge with R. anatipestifer serotype 1 strain WJ4, indicating that the mutant strain RA Delta 604 could be used as a potential vaccine candidate in the future.

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