福氏2a志贺氏菌2457T株argT基因突变体的构建

Construction of argT Gene Mutants of Shigella flexneri 2a Strain 2457T

目的:构建福氏2a志贺氏菌2457T株argT基因缺失突变体和ArgT蛋白非降解突变体,以进行后续ArgT功能研究。方法:根据福氏2a志贺氏菌2457T株基因组全序列,采用λ-Red重组系统对argT基因进行缺失,并经PCR验证;采用定点突变的方法构建ArgT非降解株,并经SDS-PAGE验证;对野生株、argT缺失突变株和ArgT非降解突变株37℃时的生长曲线及生化反应进行比较研究。结果:构建了2457T的argT缺失突变株和ArgT非降解突变株;2种突变株初始生长均较慢,但最终和野生株状态一致;2种突变株利用甘露醇的能力都比野生株强,而利用葡萄糖的能力降低。结论:获得了福氏2a志贺氏菌2457T株argT基因缺失突变体和ArgT蛋白非降解突变体。

Objective：To construc an argT deletion mutant and an ArgT non-proteolysis mutant of Shigella flexneri 2a strain 2457T for exploring the function of ArgT in S.flexneri 2a strain 2457T.Methods：According to the complete genome sequence of S.flexneri 2a strain 2457T,the argT gene was deleted by using λ-Red recombination system,and confirmed by PCR.The ArgT non-proteolysis mutant was constructed by using site-directed mutagenesis method,and confirmed by SDS-PAGE.Growth of the two mutants and wild strain at 37℃ were measured respectively,and some biochemical events of them were comparatively investigated.Results：The argT deletion mutant and ArgT non-proteolysis mutant of S.flexneri 2a strain 2457T were constructed.The two mutants grew slower than wild strain at the initial period,and ultimately they showed the consistent growth state.The capabilities to utilizing mannitol of the mutants were more than wild strain,but that to utilizing glucose were less.Conclusion：The argT deletion mutant and ArgT non-proteolysis mutant of S.flexneri 2a strain 2457T were obtained.