猪圆环病毒2型Cap全基因的克隆与原核表达  

Cloning and Prokaryotic Expression of Capsid Protein Gene from Porcine Circovirus Type 2

作　　者：李海花[1] 覃尧[2] 张全红[3] 李秀丽[1] 

机构地区：[1]天津市畜牧兽医研究所,天津300384 [2]中国农业大学动物医学院,北京100193 [3]国家知识产权局专利局专利审查协作北京中心,北京100190

出　　处：《中国畜牧兽医》2015年第2期337-341,共5页China Animal Husbandry & Veterinary Medicine

基　　金：天津市农业科学院院长基金(13004)

摘　　要：Cap蛋白是猪圆环病毒2型(porcine circovirus type 2,PCV2)的主要结构蛋白,能诱导免疫保护,是临床上利用血清学诊断PCV2的主要依据。本研究根据PCV2TJ株全基因核苷酸序列(GenBank登录号:KC751546)设计特异性引物,利用PCR从PCV2TJ株扩增获得Cap全基因,将该基因片段连接到原核表达载体pET-32a(+)中,获得重组质粒pET32a-Cap,经IPTG诱导获得约48ku的重组融合蛋白,Western blotting分析结果表明其与小鼠抗6×His单克隆抗体和PCV2阳性猪血清均呈阳性反应。本研究成功克隆Cap全基因,构建原核表达载体,并实现Cap融合蛋白的表达,这为PCV2Cap蛋白的功能研究及诊断方法的建立和亚单位疫苗的研制奠定了基础。Capsid （Cap） protein was the main structural protein of porcine circovirus type 2 （PCV2） ,and used as a main basis for PCV2 serological diagnosis in the clinical. In this study,the complete gene encoding Cap protein of PCV2 TJ strain was amplified by PCR using the specific primers designed according to the strain sequence （GenBank accession No..KC751546）,and then the amplified products were cloned into pET-32a （＋） vector to construct recombinant plasmid pET32a-Cap. The recombinant fusion protein with about 48 ku was expressed by IPTG induction. The result of Western blotting analysis showed that the fusion protein had the positive reaction with anti-6 × His tag monoclonal antibody and porcine serum against PCV2. We had successfully cloned Cap gene, constructed the prokaryotic expression vector, and then expressed Cap fusion protein,it provided the support for the establishment of PCV2 detection and development of subunit vaccine,and also made a foundation for the study of the function of PCV2.

关 键 词：猪圆环病毒2型 Cap基因 克隆 原核表达 

分 类 号：Q786[生物学—分子生物学]