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将人β-珠蛋白基因(3.1kb)反向克隆入反转录病毒载体N2A,并在N2A的LTR插入36bp的增强子HS2(简称N2A.β.E36)。将此反转录病毒重组体转染ψ-2和PA317包装细胞系,经G418筛选后,分离出完整整合的并能产生较高病毒滴度的产病毒细胞系。利用产病毒细胞系,将含增强子人β-珠蛋白基因转入小鼠胎肝细胞,Southern印迹杂交表明人β-珠蛋白基因可以完整地整合到靶细胞的基因组上,Northern杂交显示人β-珠蛋白基因可在小鼠胎肝细胞中表达。
The human β-globin gene (3.1 kb) was reverse cloned into the retroviral vector N2A and 36 bp of enhancer HS2 (N2A.β.E36) was inserted into the LTR of N2A. This retrovirus recombinant was transfected into the [psi] -2 and PA317 packaging cell lines and, following G418 selection, isolated fully virus-producing cell lines producing higher virus titers were isolated. Transfection of the human β-globin gene containing enhancer into mouse fetal hepatocytes using virus-producing cell lines showed that the human β-globin gene was completely integrated into the genome of the target cell by Southern blotting. Northern blotting showed that human β The globin gene can be expressed in mouse fetal hepatocytes.