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外源基因在受体细胞中的整合和定位,是影响体细胞基因治疗最终结果的重要环节。本实验以逆转录病毒为载体将人正常β-珠蛋白基因转移到小鼠红白血病细胞(MEL,即Friend细胞)中,探索外源基因在靶细胞中的整合、定位及其规律和不同传代对整合稳定性的影响。取外源基因转移后并经G_(418)筛选所获得的阳性克隆群体细胞进行有无G_(418)维持水平选择及不同传代数目培养,以MEL细胞为阴性对照,重组病毒(N_2HNβ)为阳性对照,提取基因组DNA。Southern
The integration and localization of exogenous genes in the recipient cells is an important part of the final result of somatic gene therapy. In this experiment, retrovirus was used as a vector to transfer human normal β-globin gene into mouse erythroleukemia cells (MEL, ie, Friend cells) to explore the integration, localization and regulation of exogenous genes in target cells and their different passage Impact on the stability of integration. The positive clonal population obtained after exogenous gene transfer and G_ (418) screening was used to select the maintenance level of G_ (418) and culture at different passage numbers. MEL cells were used as negative control and recombinant virus (N_2HNβ) was positive Control, extraction of genomic DNA. Southern