牛瑟氏泰勒虫18S rRNA基因的克隆及分子分类学分析  

Cloning and Molecular Taxonomy Analysis of 18S rRNA Gene of Cattle Theileria sergenti

作　　者：沈岩[1] 许应天[1] 李静[1] 栾杨[1] 杨兴[1] 

机构地区：[1]延边大学农学院,吉林龙井133400

出　　处：《安徽农业科学》2009年第31期15175-15176,15182,共3页Journal of Anhui Agricultural Sciences

基　　金：吉林省科技发展计划项目(20050703-4)

摘　　要：为分析牛瑟氏泰勒虫延边株18S rRNA基因序列,根据GenBank上已发表的牛瑟氏泰勒虫18S rRNA基因序列设计2对特异性引物,通过PCR扩增目的基因片段,将扩增产物连接到pMD18-T载体中,经酶切鉴定和PCR鉴定为阳性的重组质粒进行测序,对测序结果用DNAMAN软件对其与GenBank上8个虫株相关序列进行同源性比较,建立系统发育树。结果显示,牛瑟氏泰勒虫中国延边株的18SrRNA基因大小为1 744 bp,瑟氏泰勒虫延边株、兰州株、日本株以及水牛泰勒虫中国株彼此之间亲缘关系最近;瑟氏泰勒虫延边株与突变泰勒虫亲缘关系较远。Tn analyze 18S rRNA nucleotide sequence of cattle Ttwileria sergenti, two pairs of specific primers were deigned according to 18S rRNA gene of cattle Ttwileria sergenti sequences published on GenBank. 18S rRNA gene was amplified by PCR and clned into pMD18-T vector. Positive clones were identified by PCR screening and restriction digestion enzyme. The phylogenetic tree was inferred based on 18S rRNA sequence of Yanbian and the other eight species of Theileria available on GenBank. Sequencing of positive clones showed that the cloned gene has a total length of 1 744 bp. The phylogenetic tree indicated that Yanbian strain, Lanzhou strain, Japan strain of cattle T. sergenti were closer to China strain of T. buffili,but Yanbian strain was far from T. mutans.

关 键 词：牛瑟氏泰勒虫 18S RRNA基因 克隆 分子分类学 

分 类 号：S188[农业科学—农业基础科学]