肠出血性大肠杆菌O157∶H7 z0986-z1444双缺失突变株构建  

Construction of z0986-z1444 double deletion mutant of enterohemorrhagic Escherichia coli O157∶H7

作　　者：苏莉[1,2] 王建新[2] 陈芳红[2] 李崭[2] 黄洁[2] 李涛[2] 王慧[2] 

机构地区：[1]安徽医科大学,合肥230032 [2]军事医学科学院微生物流行病研究所病原微生物与生物安全国家重点实验室微生物组学与生物信息学研究室,北京100071

出　　处：《安徽医科大学学报》2017年第2期155-159,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81401643);国家重点基础研究计划(编号:2015CB554202)

摘　　要：目的采用Red重组方法敲除肠出血性大肠杆菌(EHEC)O157∶H7 z0986和z1444基因,构建双缺失突变株。方法首先构建z0986、z1444打靶片段;其次以电击法转化感受态细胞,之后通过PCR鉴定所得菌株;最终所得阳性单克隆测定生长曲线。结果 z1444单基因缺失突变株(Δz1444/933)及z0986-z1444双基因缺失突变株(Δz0986/Δz1444/933)构建成功,其生长曲线与野生型EHEC O157∶H7差异均无统计学意义。结论 z0986与z1444基因缺失对EHEC的生长无影响。构建Δz1444/933及Δz0986/Δz1444/933为深入研究z1444基因的功能及作用机制奠定了基础。Objective To construct the double deletion mutant of enterohemorrhagic Escherichia coli（EHEC） O157 ： H7 by knocking out z0986 gene and z1444 gene with Red recombinant system. Methods Firstly, the target fragments of z0986 and z1444 were established. Secondly, competent cells were struck by lightning, then the bacteria were identified through PCR. The growth curve of the ultimate positive monoclone was assayed. Results The solo deletion mutant（Az1444/933） and the double deletion mutant（AzO986/Az1444/933） were successfully construc- ted and their growth curves had no significant difference compared to the wild type EHEC. Conclusion The deletion mutant of z0986 and z1444 has no effect on growth of EHEC. The construction of Az1444/933 as well as Az0986/Az1444/933 would be helpful for further study of functions and mechanisms of z1444 gene.

关 键 词：肠出血性大肠杆菌 RED重组系统 三型分泌系统 

分 类 号：Q939.93[生物学—微生物学]