副猪嗜血杆菌EZ-Tn5转座子插入突变体库的构建及减毒株的筛选

Construction of Mutation Library in Haemophilus parasuis by Inserting Tn5 Transposon and the Screening of Attenuated Strain

[目的]获得副猪嗜血杆菌减毒株。[方法]应用转座子技术构建转座子插入突变体库,卡那抗性筛选阳性菌株,PCR扩增卡那特异片段去除假阳性,小鼠感染试验检测突变株毒力,并对获得的减毒株进行生物学特性检测。[结果]所获得减毒突变菌株具有与野毒株相似的增殖能力,传代后毒力稳定,遗传学特性稳定。[结论]该研究结果为进一步探讨HPS毒力因子、致病机制奠定了基础。

[Objective]The study aimed to obtain attenuated strain of Haemophilus parasuis.[Method]Tn5 transposon technology was used to construct a library of mutants.Positive mutants were screened by kanamycin resistance.False positive were identified by PCR and then removed.Mice were infected to detect the virulence of mutants.The bionomics of attenuated strains were detected,too.[Result]The attenuated mutants showed similar reproductive activity to that of wild strain.The virulence of mutants was stable after 30 passages.[Conclusion]This study provided foundation for exploring the virulence factors and pathogenic mechanism of HPS.