乙肝病毒e抗原反式激活蛋白的亚细胞定位研究

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DEGFP-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。

Subcellular location of the HBeAg transactivated protein

Objective To prove that HBeAg transactivated protein can express proteins in cells and find the subcellular location of the proteins expressed by HBeAgTP.Methods pGBKT7-HBeAgTP and green fluorescent protein(GFP)expression vector pEGFP-HBeAgTP was established.The pEGFP-HBeAgTP was transfected in HepG2 cells,and the subcellular location of the proteins expressed by HBeAgTP analyzed through green fluorescent microscopy after 24 hours.The pGBKT7-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.Conclusion HBeAgTP can express protein,and the protein expressed by HBeAgTP subcellularly located in cytoplasm.