摘要
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.
出处
《武汉大学学报（理学版）》
CAS
CSCD
北大核心
2001年第6期732-736,共5页
Journal of Wuhan University:Natural Science Edition
基金
国家 8 6 3高技术新探索项目 ( 39880 0 31)
