新城疫病毒B_(95)株HN蛋白基因的克隆及序列分析  

Cloning and sequence analysis of HN gene of NDV B_(95)strain

作　　者：田兴华[1] 陈燕军[2] 郝勤宗[1] 裘孝良[1] 

机构地区：[1]河北农业大学动物科技学院,河北保定071001 [2]中国农科院上海家畜寄生虫病研究所,上海徐汇200232

出　　处：《中国兽医杂志》2003年第10期7-10,共4页Chinese Journal of Veterinary Medicine

基　　金：上海市科技兴农重点攻关项目 (农科攻字 2 0 0 0 ;第 5 -0 1号 ) ;河北省自然科学基金项目 (3 980 84)。

摘　　要：根据 NDV　 HN蛋白已知基因序列设计一对引物 ,以提取的澳大利亚布里斯班 NDV分离株 B95基因组 RNA为模板 ,利用 RT- PCR技术扩增得到一条约 1.9kb的条带。将扩增产物克隆到 PGEM- T- easy载体中 ,并对重组质粒进行酶切分析、PCR鉴定 ,结果证明我们得到了 HN基因的阳性重组子 ,随后采用 Sanger's双脱氧末端终止法对阳性重组子进行核苷酸序列测定 ,获得该基因全长序列 ,并进行同源性分析。 B95株与其他毒株核苷酸同源性为 95 .1%～ 87% ,氨基酸同源性为96 .1%～ 92 .1%。According to published HN protein gene sequence of NDV, we designed and synthesiezed a pair of primers. Genetic RNA of NDV B (95)strain, which was isolated from Brisbane in Australia, was used as template from cDNA synthesis of HN gene. A 1.9 Kb fragment was amplified from B (95) strain and cloned into the PGEM-T-easy vector. The recombinant plasmid was proved to be true by enzyme and PCR. The nucleotide sequence of HN gene was obtained by sanger's sequencing technique. Compared HN gene sequence of B (95)strain with the corresponding sequence of other strains of NDV, the homology of the nucleotide sequence was 95.1%～87%, the homology of the protein sequence was 96.1%～92.1%.

关 键 词：新城疫病毒 B95株 HN蛋白基因 克隆 序列分析 NDV RT—PCR 重组质粒 氨基酸 同源性分析 核苷酸 

分 类 号：S852.659.5[农业科学—基础兽医学] Q785[农业科学—兽医学]