摘要
目的:构建人bax真核表达载体,经脂质体导入人胃癌耐药细胞SGC7901/VCR,并检测bax基因在转导株的表达.方法:采用分子克隆技术将bax基因插入真核表达载体pBK-CMV的多克隆位点之间,以脂质体介导法将目的基因导入受体细胞SGC7901/VCR,G418筛选克隆细胞,Westernblot检测bax基因的表达.结果:成功地构建了重组质粒pBK-bax,将之转入细胞后,经G418筛选30d获得了稳定的细胞克隆,即SGC7901/VCR-baxcels.Western印迹证实了bax基因转导株具有明显的Bax蛋白表达.结论:bax真核表达载体的构建,为胃癌耐药的临床治疗打下了基础.
AIM: To construct a human bax eukaryotic expression vector and to detect its expression in human gastric cancer drug resistant SGC7901/VCR cells. HZMETHODS: bax gene was first inserted into polyclonal sites of pBKCMV vector, followed by the transfection of recombinant plasmid into vincristine resistant human gastric cancer SGC7901/VCR cells by lipofectamine. G418 was used to select resistant clones. Then, Western blot was used to detect the expression of Bax protein. RESULTS: Recombinant plasmids were successfully constructed. After they were transfected into human gastric cancer drug resistant SGC7901/VCR cells, stably transfected cell clones were obtained 30 d after G418 selection. Western blot indicated obvious expression of Bax protein in bax transfected cells. CONCLUSION: The establishment of bax gene stable transfectant provides a sound basis for further investigation of gastric cancer multidrug resistance.
出处
《第四军医大学学报》
1999年第6期468-471,共4页
Journal of the Fourth Military Medical University