蓝舌病毒1型VP2基因在昆虫杆状病毒表达系统中的表达  

Expression of VP2 gene of bluetongue virus type 1 in the baculovirus expression system

作　　者：李扬帆[1] 独军政[1] 邵军军[1] 高闪电[1] 林彤[1] 赵付荣[1] 常惠芸[1] 

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室国家口蹄疫参考实验室,甘肃兰州730046

出　　处：《中国兽医科学》2015年第3期265-269,共5页Chinese Veterinary Science

基　　金：国家生猪产业体系项目(CARS-36-068)

摘　　要：为了研制蓝舌病新型亚单位疫苗,将蓝舌病毒1型(BTV1)VP2基因克隆到pFastBacHTB载体中,得到重组转座载体pVP2,并转化至DH10Bac感受态细胞,获得了重组杆粒rVP2。然后采用脂质体转染法转染Sf9昆虫细胞,通过克隆筛选获得重组杆状病毒。SDS-PAGE分析结果显示,获得了约110ku的表达产物,与预期结果相符。Western-blot结果显示,目的蛋白能被BTV1感染的山羊阳性血清识别。结果表明,经杆状病毒体系表达的可溶性重组VP2蛋白具有良好的反应原性,为进行蓝舌病新型亚单位疫苗的研制奠定了基础。In order to develop new subunit vaccines against bluetongue,the VP2 gene from bluetongue virus type 1(BTV1)strain was cloned into the baculovirus expression vector pFastBacHTB and the recombinant plasmid pVP2 was obtained.The recombinant plasmid pVP2 was transformed into DH10 Bac competent cells.To get the recombinant baculovirus,the recombinant bacmid rVP2 was transfected into Sf9 cells by liposome.Expression of VP2 was identified by SDS-PAGE.In result,the expressed VP2 protein is 110 ku approximately,which correspond to the anticipated size.The expressed VP2 protein can specifically react with BTV1 positive serum.The result showed that the soluble VP2 protein expressed in the baculovirus expression system had good reactionogenicity.It laid the foundation for development of new subunit vaccines against bluetongue.

关 键 词：蓝舌病毒1型 VP2基因 杆状病毒 表达 

分 类 号：S852.65[农业科学—基础兽医学]