Efficient construction of Haemophilus parasuis mutants based on natural transformation

文献类型： 外文期刊

第一作者： Li, Junxing

作者： Li, Junxing;Yuan, Xiufang;Xu, Lihua;Kang, Lei;Jiang, Jun;Wang, Yicheng

作者机构：

期刊名称：CANADIAN JOURNAL OF VETERINARY RESEARCH-REVUE CANADIENNE DE RECHERCHE VETERINAIRE （ 影响因子：1.31； 五年影响因子：1.438 ）

ISSN： 0830-9000

年卷期： 2016 年 80 卷 4 期

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收录情况： SCI

摘要： Studies on virulence factors and pathogenecity of Haemophilus parasuis have long been hindered by a lack of a consistent system for genetic manipulation. In this study, competence was induced by transferring H. parasuis from rich medium to starvation medium media-IV (M-IV) and iscR gene deficient mutants of H. parasuis were generated efficiently. Transformation frequency varied from 4.1 x 10(-5) to 1.1 x 10(-8) when using circular plasmid, and increased to about 2- to 31-fold when transformed using linearized plasmid. Allele replacement occurred efficiently in 6 strains, which are transformable using both circular and linearized pTRU, but not in another 2 strains which could only be transformed using linearized plasmid. The iscR mutants were stable for at least 20 passages in vitro. Haemophilus parasuis strains vary extensively in natural transformation efficiency and the method established here allows for transformation of a larger spectrum of strains with an easily accessed plasmid. This provides important tools for genetic manipulation of H. parasuis.

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