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重组人凋亡诱导因子基因的构建、表达及其对HeLa细胞的促凋亡作用
引用本文:于翠娟,孟艳玲,桂俊豪,赵晶,金明,王智,王成济,杨安钢.重组人凋亡诱导因子基因的构建、表达及其对HeLa细胞的促凋亡作用[J].生物化学与生物物理进展,2002,29(6):915-921.
作者姓名:于翠娟  孟艳玲  桂俊豪  赵晶  金明  王智  王成济  杨安钢
作者单位:第四军医大学生物化学与分子生物学教研室,西安,710033
基金项目:国家杰出青年科学基金资助项目(39925036), 军队杰出中青年人才科研基金资助项目(98J009)和国家高技术“863”计划资助项目(2001AA217101).
摘    要:凋亡诱导因子(apoptosis-inducing factor, AIF)定位于细胞的线粒体膜间隙.当凋亡信号刺激时,AIF分子从线粒体释放到胞浆,然后转位到细胞核内,引起染色体核周边凝集和DNA呈大片段断裂(~50 kb).用RT-PCR法分段克隆得到人全长AIF基因,经改造截去其N端线粒体定位信号编码序列,代之以不同长度的绿脓杆菌外毒素(PE)转膜结构域序列.把这些重组基因克隆入pIRES2-EGFP真核表达载体,脂质体法转染HeLa细胞,通过荧光显微镜观察、共聚焦显微镜观察、电镜观察等方法检测了多种重组人AIF基因的表达及其对细胞生长的影响.证明了重组人AIF基因的表达可引起HeLa细胞死亡,为肿瘤的杀伤提供了新的策略.

关 键 词:AIF,细胞凋亡,融合蛋白,HeLa细胞
收稿时间:2002/4/21 0:00:00
修稿时间:2002年4月21日

Gene Construction, Expression and The Apoptosis Inducing Effects on HeLa Cells of The Recombinant Apoptosis Inducing Factor
YU Cui-Juan,MENG Yan-Ling,GUI Jun-Hao,ZHAO Jing,JIN Ming,WANG Zhi,WANG Cheng-Ji and YANG An-Gang.Gene Construction, Expression and The Apoptosis Inducing Effects on HeLa Cells of The Recombinant Apoptosis Inducing Factor[J].Progress In Biochemistry and Biophysics,2002,29(6):915-921.
Authors:YU Cui-Juan  MENG Yan-Ling  GUI Jun-Hao  ZHAO Jing  JIN Ming  WANG Zhi  WANG Cheng-Ji and YANG An-Gang
Institution:Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China;Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China;Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China;Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China;Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China;Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China;Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China;Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China
Abstract:
Keywords:apoptosis-inducing factor (AIF)  apoptosis  fusion protein  HeLa cell
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