抗新城疫病毒HN蛋白单抗可变区基因的克隆与序列分析  

Clone and Sequence Analysis of Variable Region Gene of a McAb Against NDV HN Protein

作　　者：路希山[1] 黄艳艳[2] 胡北侠[2] 李士娟 于周[1] 李相钊[1] 王金宝[4] 张秀美[2] 

机构地区：[1]青岛农业大学动物科技学院,青岛266109 [2]山东省农业科学院畜牧兽医研究所,济南250000 [3]济南市农业开发办公室,济南250000 [4]山东省农业科学院,济南250000

出　　处：《中国畜牧兽医》2008年第5期25-28,共4页China Animal Husbandry & Veterinary Medicine

基　　金：山东省科技攻关项目(022020103-02)

摘　　要：提取抗新城疫HN蛋白单克隆抗体C3-B7杂交瘤细胞总RNA,进行RT-PCR,扩增出轻重链可变区基因。凝胶回收纯化后与pMD-18T载体连接,重组载体转化于宿主菌DH5α,筛选出阳性重组子,菌液PCR鉴定后进行测序。结果显示,VH基因全长357 bp,编码119个氨基酸;VL基因全长332 bp,编码110个氨基酸。这为单链抗体的构建及表达奠定了基础。With the total RNA from C3-B7 hybridoma cells which secrete monoclonal antibody against NDV HN protein, the VH（heavy-chain variable region） and VL（light-chain variable region）genes encoding the monoclonal antibody were ampli fled by RT-PCR. The purified pruduct was cloned to pMD18-T vector and the recombined plasmids were transformed into DH5a. Colonies were selected and the specific recombinant plasmid was identified by colony PCR. The result shows that the VH gene encoding 119 aa and the VL gene encoding 110 aa were 357 bp and 332 bp, respectively. This establish the foundation for the construction and expression of single chain Fv（svFv）.

关 键 词：新城疫病毒 HN蛋白 可变区基因 克隆 序列分析 

分 类 号：S852.659.5[农业科学—基础兽医学]