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）.
出处
《中国畜牧兽医》
CAS
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)
关键词
新城疫病毒
HN蛋白
可变区基因
克隆
序列分析
newcastle disease virus
HN protein
variable region gene
clone
sequence analyse
