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用重组人β-珠蛋白基因的逆转录病毒载体转染单向性的ψ-2包装细胞系,继而感染双向性的PA317包装细胞系,分离到能产生重组逆转录病毒载体的细胞系PIWβ,其分泌的重组载体滴度达1.1×10~5CFU/ml。利用这种缺失性病毒粒子,将人β-珠蛋白基因高效地转移到Friend细胞,Southern印迹杂交结果证实,人β-珠蛋白基因可以完整、稳定地整合到靶细胞的基因组上。以上结果为人β-地中海贫血的基因治疗研究提供了有用的参考途径。
Transfection of the uniportabilized ψ-2 packaging cell line with a retroviral vector of recombinant human β-globin gene followed by infectivity of the bi-directional PA317 packaging cell line and isolation of a cell line capable of producing a recombinant retroviral vector, PIWβ, The secreted recombinant vector titer reached 1.1 × 10 ~ 5CFU / ml. Using this deletional virion to efficiently transfer human β-globin gene to Friend cells, Southern blot hybridization results confirm that the human β-globin gene can be integrated intact and stably into the target cell’s genome. The above results provide a useful reference for the study of gene therapy of human β-thalassemia.