猪瘟病毒E_2(gp55)基因在昆虫细胞中的表达  

Expression of E_2 gene of classical swine fever virus in insect cell

作　　者：季新成[1] 陈杰[2] 张彦明[1] 吴发兴[2] 黄宝续[2] 李晓成[2] 张燕霞[2] 许信刚[1] 

机构地区：[1]西北农林科技大学动物科技学院,陕西杨凌712100 [2]农业部动物检疫所国家动物流行病学研究中心,山东青岛266032

出　　处：《中国预防兽医学报》2004年第4期256-258,共3页Chinese Journal of Preventive Veterinary Medicine

基　　金：陕西省科技攻关项目 (2 0 0 2K0 2 -G8- 0 3:2 0 0 3K0 2 -G11- 0 1) ;陕西省农业分子生物学重点实验室项目 (2 0 0 2 - 6 ) ;国家流行病学专题项目 (CECR - 2 )

摘　　要：将除去 3’端跨膜区的猪瘟病毒E2 基因亚克隆到杆状病毒转移载体pFastBacHTb ,获得重组质粒pFBHT_E2 ,转化进含穿梭载体Bacmid的感受态细胞DH10Bac ,发生转座作用 ,经抗性及蓝白斑筛选得到含E2 基因的重组载体质粒rBacmid_E2 ,以脂质体介导的方法将此重组载体质粒转染sf9昆虫细胞 ,获得重组病毒 ,命名为rvBac_E2 。经SDS_PAGE和Western_blot及ELISA等方法检测 ,结果表明 ,此E2 基因在昆虫细胞中正确表达 ,表达的蛋白能与猪瘟阳性血清发生特异性反应。The E_2 gene of Classical Swine Fever Virus(CSFV)was subcloned into baculovirus transfer vector and recombinant baculovirus vector pFBHT-E_2 was extracted.Then it was transferred into E.coli DH10Bac and recombinant bacmid was constructed.The recombinant baculovirus was obtained after the tranferation of the recombinant bacmid into sf_9 cell,named as rvBac-E_2.The techniques of SDS-PAGE,Westernblot and ELISA were used to detect and identify the products expressed in sf_9 cell by recombinant baculovirus.The results indicated the E_2 gene was properly expressed in sf_9 cell,and the expressed protein could be recognized by the positive serum of CSFV.

关 键 词：猪瘟病毒 E2基因 杆状病毒表达载体 表达 

分 类 号：S813.3[农业科学—畜牧学] S852.651[农业科学—畜牧兽医]