大肠杆菌O110 O-抗原基因簇序列的破译及特异基因鉴定  

Molecular characterization of O-antigen gene cluster of Escherichia coli O110 reference strain and identification of O110 specific genes

作　　者：夏秋雨[1] 王威[2] 王岩[3] 李雅玥[2] 郭宏杰[2] 王磊[2] 耿运琪[1] 

机构地区：[1]南开大学生命科学学院,天津300071 [2]南开大学泰达生物技术学院 [3]天津市第四医院

出　　处：《中国人兽共患病杂志》2005年第2期125-128,118,共5页Chinese Journal of Zoonoses

基　　金：国家"863计划"(2002AA2Z2051)

摘　　要：目的定大肠杆菌O110标准株的O-抗原基因簇序列,对其基因结构进行遗传学分析,鉴定可用于快速分型的特异DNA序列。方法用长距离PCR扩增O-抗原基因簇序列,建立鸟枪法文库进行随机测序,用生物信息学的方法进行序列拼接与分析,根据测定的序列设计引物,用PCR的方法确定针对大肠杆菌O110的特异基因。结果与结论确定了大肠杆菌O110标准株的O-抗原基因簇序列以及该基因簇中7个开放阅读框架(openreadingframe,orf)的功能,分别为糖基转移酶基因(orf1,orf2,orf7),小分子修饰酶基因(orf3,orf5),O-抗原转运酶基因(wzx)和O-抗原聚合酶基因(wzy)。鉴定出了可以用于PCR方法对大肠杆菌O110快速检测的特异基因。In the present study, the sequences of O-antigen gene cluster in the reference strain of Escherichia coli O110 were determined and the specific DNA used for rapid genotyping was identified. The chromosomal DNA was extracted from reference strain of E. coli O110 and the O-antigen gene cluster was amplified by long-range PCR. Shot-gun bank was set up and the clones were selected and sequenced later on. All readings were assembled and analyzed with bioinformatics method. Primers were designed according to the desired sequences and used in the PCR to identify the specific genes of E. coli O110. It was found that the sequences of O-antigen gene cluster E. coli O110 were identified by the methods used in present study, in which 7 genes located in O110 gene cluster were found, including 3 glycosyl transferase genes (orf1, orf2, orf7), 2 O-antigen modification genes (orf3, orf5), the O-antigen polymerase gene (wzy) and the O-antigen transferase gene (wzx). These 3 glycosyl transferase genes were identified to share high specificity with E. coli O110.

关 键 词：大肠杆菌 O-抗原基因簇 特异基因 分子分型 

分 类 号：R378.2[医药卫生—病原生物学]