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- W921644713 abstract "目的构建DJ-1 shRNA真核表达载体，获得干扰质粒稳定转染的人肝癌HepG2细胞株。方法合成DJ-1基因干扰序列，定向插入真核表达载体pGenesil-1质粒并测序鉴定；干扰质粒经脂质体介导转染人肝癌HepG2细胞，G418（450mg／L）有限稀释法持续筛选8周后，扩增获得稳定转染的细胞株；荧光显微镜观察转染扩增效率；逆转录．聚合酶链反应（RT—PCR）和Westernblot检测稳定转染细胞株的DJ-1 mRNA和蛋白的表达。结果测序证明DJ-1干扰序列及读码框完全正确，G418筛选后扩增细胞中稳定转染扩增百分率高达75．00％。稳定转染细胞DJ-1 mRNA表达下调，DJ-1蛋白抑制率为98．78％（P〈0．05）。结论成功构建DJ-1 shRNA真核表达载体，DJ-1 shRNA质粒稳定转染HepG2细胞株的建立，为以DJ-1基因为靶点的人肝癌基因研究提供实验基础。" @default.
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- W921644713 date "2011-01-01" @default.
- W921644713 modified "2023-09-24" @default.
- W921644713 title "DJ-1 shRNA表达载体的构建及其稳定转染人肝癌细胞HepG2细胞株的建立" @default.
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