用宿主范围扩大的重组AcNPV在家蚕中表达TNF

用宿主范围扩大的重组AcNPV在家蚕中表达TNF Expression of Tumor Necrosis Factor in Bombyx mori by Host Range Expanded Recombinant AcNPV 胡敏 HU Min 1 2 first-author 刘映乐 LIU Ying-le 1 2 齐义鹏 QI Yi-peng 1 2 朱应 ZHU Ying 1 2 武汉大学病毒研究所 武汉大学病毒研究所 Institute of Virology, Wuhan University Institute of Virology, Wuhan University 武汉大学病毒研究所,湖北,武汉,430072 武汉大学病毒研究所,湖北,武汉,430072 用宿主范围扩大的核型多角体病毒双穿梭载体 Bonpvid与重组转座质粒 p Fast- TNF转座 ,使含有TNF基因的表达盒插入 Bonpvid的 Tn7att的接受位点筛选阳性重组 E.coli菌落 .将提取的阳性重组子 DNA分别转染家蚕 Bm N细胞和 Sf- 9细胞 ,获得了能表达 TNF的重组病毒 .SDS- PAGE和 Western blot分析表明 ,TNF在上述两种细胞中得到表达 .以家蚕作为生物反应器 ,将含有重组病毒的细胞上清注射感染家蚕 4龄幼虫 ,使TNF在蚕体表达 .用 Bm N细胞中表达的 TNF感染肿瘤细胞 ,诱导细胞发生了凋亡 . Recombinant virus vAcBmT was obtained after transposition of pFast-TNF into Tn7 att in bonpvid which is a host range expanded Bi-shuttle vector by Bac-to-Bac system.vAcBmT DNA transfected BmN cell and Sf-9 cell respectively, SDS-PAGE and Western blot showed that the TNF was expressed in both of the cells. The viral stock of vAcBmT was then infected into Bombyx mori larvea, analysis showed that the TNF was expressed in it. After the TNF which expressed in Bombyx mori larvea was added into Hela cell, it induced the cell apoptosis. 肿瘤坏死因子（TNF）-α BactoBac系统 宿主范围扩大的重组AcNPV 家蚕 凋亡 tumor necrosis factor-α Bac-to-Bac system host range expanded recombinant AcNPV Bombyx mori apoptosis Q784 国家 8 6 3高技术新探索项目 （ 39880 0 31） 2001-06-01 2021-04-01 6