| 线粒体基质蛋白Hsp70、Hsp60的表达、纯化、鉴定及抗体制备 |
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| 引用本文： | 程晓丽,李春燕,连建华,齐华,刘永波.线粒体基质蛋白Hsp70、Hsp60的表达、纯化、鉴定及抗体制备[J].郑州大学学报(医学版),2006,41(6):1058-1060. |
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| 作者姓名： | 程晓丽 李春燕 连建华 齐华 刘永波 |
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| 作者单位： | 郑州大学基础医学院细胞生物学与医学遗传学教研室,郑州,450052;北华大学医学院附属医院手外科,吉林,132011 |
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| 基金项目： | 河南省郑州大学校科研和教改项目 |
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| 摘 要： | 目的:建立有效的线粒体基质蛋白Hsp70、Hsp60原核细胞表达系统,获得纯度较高的Hsp70、Hsp60蛋白,并制备抗体.方法:以HeLa细胞cDNA为模板扩增成熟的hsp70、hsp60 DNA,插入pPRO-EX-HTb质粒,IPTG诱导B21 E.coli表达,Ni2 离子螯合树脂纯化蛋白,获取被免疫的兔抗血清,纯化后进行Western-blot鉴定.结果与结论:成功构建Hsp70、Hsp60高效原核表达系统并获得特异性较好的抗体,为进一步研究线粒体基质蛋白的功能奠定了基础.
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| 关 键 词： | 线粒体基质蛋白 热休克蛋白 表达 纯化 Western-blot分析 抗体 |
| 收稿时间： | 2006-09-19 |
| 修稿时间： | 2006年9月19日 |
Expression and identification of mitochondrial heat shock protein 70, 60, and its antibody |
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| CHENG Xiaoli,LI Chunyan,LIAN Jianhua,QI Hua,LIU Yongbo.Expression and identification of mitochondrial heat shock protein 70, 60, and its antibody[J].Journal of Zhengzhou University: Med Sci,2006,41(6):1058-1060. |
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| Authors: | CHENG Xiaoli LI Chunyan LIAN Jianhua QI Hua LIU Yongbo |
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| Institution: | 1. Department of Cell Biology and Medical Genetics, College of Basic Medical Sciences, Zhengzhou University, Zhengzhou 450052 ;2.Department of Hand Surgery, North Huada Medical College Accessorial Hos- pital, Jilin 132011 |
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| Abstract: | Aim: To get efficient expression vector of mitochondrial matrix protein 70 and 60.Methods:The full-length mitochondrial heat shock protein 70 and 60 (Hsp70 and Hsp60) DNA was amplified from a cDNA library of human HeLa cells. E.coli B21 was transformed by pPRO-EX-HTb plasmid which contained target DNA, then each protein was purified after E.coli amplifying and target DNA expressing. The serum of immunized rabbit was obtained, and Western-blot analysis was done after being purified.Results and Conclusion: The pPRO-EX-HTb expression vectors have been constructed successfully. And antibodies have been acquired, which have laid the foundation for further research on the function of mitochondrial matrix protein. |
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| Keywords: | mitochondrial matrix protein heat shock protein expression purify Western-blot analysis antibody |
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