弗氏2a志贺菌htrA基因突变体的构建

Construction of HtrA Mutants of Shigella flexneri

目的:构建弗氏2a志贺菌2457T的htrA基因缺失突变株及HtrA酶失活突变株,以便进一步研究HtrA蛋白的功能。方法:用PCR扩增htrA基因上下游同源臂,构建含有kan基因的打靶片段,采用λ-Red重组系统对htrA基因进行缺失,用PCR进行验证;通过定点突变的方法构建HtrA酶失活突变株,并测序验证。结果与结论:构建了2457T htrA缺失突变株和2457T/htrAFSA酶失活突变株。

Objective： To explore the function of htrA gene by constructing an htrA deletion mutant and an HtrA enzyme-inactived mutant of Shigella flexneri 2a strain 2457T.Methods： htrA gene was deleted by using λ-Red recombination system,and confirmed by PCR.The HtrA enzyme-inactived mutant was constructed by using site-directed mutagenesis method,and confirmed by sequencing.Results Conclusion： The htrA deletion mutant and HtrA enzyme-inactived mutant of S.flexneri 2a strain 2457T were constructed.