番鸭细小病毒NS1基因的克隆与原核表达  

CLONING AND PROKARYOTIC EXPRESSION OF NS1 GENE OF MUSCOVEY DUCK PARVOVIRUS

作　　者：董嘉文[1] 孙敏华[1] 李林林[1] 袁建丰[1] 邝瑞欢[1] 胡奇林[1] 

机构地区：[1]广东省农业科学院动物卫生研究所广东省兽医公共卫生公共实验室,广州510640

出　　处：《中国动物传染病学报》2013年第3期33-37,共5页Chinese Journal of Animal Infectious Diseases

基　　金：公益性行业(农业)科研专项(201003012);广东省农业科学院动物卫生研究所所长基金

摘　　要：本研究利用PCR方法扩增番鸭细小病毒(Muscovy duck parvovirus,MPV)非结构蛋白NS1基因,并将其克隆至原核表达载体pET32a(+),构建重组表达质粒pET32a-NS1。将其转化受体菌E.coli Rosetta感受态细胞,经IPTG诱导表达。SDS-PAGE和Western blot试验结果表明,NS1基因在大肠杆菌中得到了表达,并且可与抗MPV的多克隆抗体发生特异性反应。MPV NS1基因的成功表达为研制MPV的诊断试剂盒、基因工程疫苗等奠定了基础。The NS1 gene of Muscovy duck parvovirus（MPV） was amplified by PCR and inserted into multiple clone sites of pET32a（＋） vector.The insertion of NS1 gene was confirmed in restriction endonuclease analysis and sequencing.The recombinant plasmid was transformed into E.coli Rosetta and NS1protein was expressed with induction of 1.0 mmol/ L IPTG.The recombinant NS1 protein was visualized in SDS-PAGE and recognized by MPV specific antibody in Western blot.The successful expression of NS1 gene laid the foundation for development of diagnosis reagent kit and genetically engineering vaccine.

关 键 词：番鸭细小病毒 NS1基因 克隆 表达 

分 类 号：S852.659.2[农业科学—基础兽医学]