乙肝病毒e抗原反式激活蛋白的亚细胞定位研究  

Subcellular location of the HBeAg transactivated protein

作　　者：王建军[1] 赵平[1] 靳雪源[1] 成军[2] 卿松[1] 赵志海[1] 丁宁[1] 

机构地区：[1]解放军第三○二医院,北京100039 [2]北京市地坛医院传染病研究所

出　　处：《中华实验和临床病毒学杂志》2006年第4期393-395,共3页Chinese Journal of Experimental and Clinical Virology

摘　　要：目的验证HBeAgTP基因在细胞内的表达及表达蛋白的亚细胞定位。方法构建HBeAgTP基因的酵母表达诱饵质粒pGBKT7-HBeAgTP及绿色荧光蛋白表达质粒pEGFP-C1-HBeAgTP,pEGFP-C1-HBeAgTP转染HepG2细胞,24h后荧光显微镜下观察蛋白表达的亚细胞定位。pGBKT7-HBeAgTP转化AH109酵母细胞,提取转化了质粒的酵母蛋白质,进行Western免疫印迹分析。结果成功构建出HBeAgTP基因。HBeAgTP基因的酵母表达诱饵质粒pGBKT7-HBeAgTP及绿色荧光蛋白表达质粒pEGFP-C1-HBeAgTP,Western免疫印迹法印证HBeAgTP可表达蛋白,其表达的蛋白亚细胞定位于细胞质。结论HBeAgTP基因可表达蛋白,其表达蛋白亚细胞定位于细胞质。Objective To prove that HBeAg transactivated protein can express proteins in cells and find the subeellular location of the proteins expressed by HBeAgTP. Methods pGBKTT-HBeAgTP and green fluorescent protein （GFP） expression vector pEGFP-HBeAgTP was established. The pEGFP-HBeAgTP was transfected in HepG2 cells, and the subeellular location of the proteins expressed by HBeAgTP analyzed through green fluorescent microscopy after 24 hours. The pGBKTT-HBeAgTP was transformed into yeast AH109, and Western Blotting was employed to analyze the yeast protein transformed with the vector. Results pGBKT7- HBeAgTP and pEGFP-HBeAgTP gene was successfully cloned, HBeAgTP could express protein in cells proved by Western Blotting, and the protein expressed by HBeAgTP subcellularly located in cell plasma. Condusion HBeAgTP can express protein, and the protein expressed by HBeAgTP subcellularly located in cytoplasm.

关 键 词：肝炎E抗原 乙型 免疫印迹法 

分 类 号：R392[医药卫生—免疫学]