截短型人凋亡诱导因子AIF△1-400对HeLa细胞的促凋亡作用

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目的:观察人截短型AIF(AIF△1-400)对HeLa细胞的促凋亡作用.方法:在AIF△1-120基因克隆成功的基础上进一步改造,构建截去AIF基因线粒体定位信号、FAD结合结构域和NADH结合结构域(1~400位氨基酸)编码序列的AIF△1-400基因,将其克隆入pcDNA3真核表达载体,用脂质体法转染HeLa细胞,通过Western blot检测该基因在转染细胞中的表达通过电镜观察截短型分子对肿瘤细胞的具有促凋亡活性.结果:经酶切鉴定与测序证实,AIF△1-400的真核表达载体构建成功.通过Western blot证实截短型基因在HeLa细胞中表达,电镜观察可见转染细胞骨架的破坏和细胞核固缩等凋亡特征.结论:人截短型AIF(AIF△1-400)基因的表达可诱导HeLa细胞凋亡.
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