| 分子伴侣BiP结构域分析及对肝癌细胞增殖和凋亡的影响 |
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| 引用本文： | 李祥柱,刘艳娜,赵文君,周菁华,郭风劲.分子伴侣BiP结构域分析及对肝癌细胞增殖和凋亡的影响[J].细胞生物学杂志,2011(11). |
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| 作者姓名： | 李祥柱 刘艳娜 赵文君 周菁华 郭风劲 |
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| 作者单位： | 重庆医科大学细胞生物学及遗传学教研室;重庆医科大学分子医学及肿瘤研究中心; |
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| 基金项目： | 国家自然基金(No.31040019); 教育部留学人员基金(No.2009-1590)资助项目~~ |
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| 摘 要： | 结合免疫球蛋白(binding immunoglobulin protein,BiP),是Hsp70(70 kilodalton heatshock proteins)蛋白家族的成员之一,是内质网应激(endoplasmic reticulum stress,ERS)主要的调节器。为了研究BiP的分子结构与生物学功能的关系,首先,分析BiP的DNA序列和蛋白质空间构象,接着利用重叠PCR方法分别克隆ATPase结构域缺失体和peptide-binding结构域缺失体(BiPa和BiPp),成功构建带myc标签的真核表达载体,转染LO2细胞和SMMC-7721细胞,应用免疫印迹方法检测其在细胞内的表达;运用MTT法、BrdU免疫组化法及流式细胞仪分别检测细胞的增殖和凋亡;检测BiP及两个组成结构域对SMMC-7721细胞增殖和凋亡的影响。酶切、电泳和DNA测序结果显示,BiP全长和两个缺失体的真核表达载体构建成功;免疫印迹检测到BiP三种真核载体在LO2细胞和SMMC-7721细胞中均能正确表达;MTT和BrdU免疫组化结果表明,BiP全长和缺失体BiPa、BiPp三种真核载体均能有效促进SMMC-7721的增...
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| 关 键 词： | BiP 缺失体 生物信息学 细胞增殖 |
Analysis on Molecular Chaperone BiP Domain and Influence on Proliferation and Apoptosis in Hepatoma Cells |
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| Li Xiangzhu,Liu Yanna,Zhao Wenjun,Zhou Jinghua,Guo Fengjin.Analysis on Molecular Chaperone BiP Domain and Influence on Proliferation and Apoptosis in Hepatoma Cells[J].Chinese Journal of Cell Biology,2011(11). |
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| Authors: | Li Xiangzhu Liu Yanna Zhao Wenjun Zhou Jinghua Guo Fengjin |
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| Institution: | Li Xiangzhu,Liu Yanna,Zhao Wenjun,Zhou Jinghua,Guo Fengjin* (Department of Cell Biology and Genetics,Chongqing Medical University,Molecular Medicine and Cancer Research Center,Chongqing 400016,China) |
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| Abstract: | Binding immunoglobulin protein(BiP) is a resident member of Hsp70 family and master regulator for Endoplasmic Reticulum Stress(ERS).To study the relationship of Bip domain with the physiological function,BiP DNA sequence and protein structure were analysed,two deletion mutants of BiP and BiP full-length eukaryotic expression vectors with myc tag were successfully constructed with the method of overlapping PCR mutagenesis respectively.BiPa deletion mutant is BiP mutant with ATPase domain deleted,BiPp deletio... |
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| Keywords: | BiP deletion bioinformatics cell proliferation |
| 本文献已被 CNKI 等数据库收录！ |
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