线粒体基质蛋白Hsp70、Hsp60的表达、纯化、鉴定及抗体制备  

Expression and identification of mitochondrial heat shock protein 70, 60, and its antibody

作　　者：程晓丽[1] 李春燕[2] 连建华[1] 齐华[1] 刘永波[1] 

机构地区：[1]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 [2]北华大学医学院附属医院手外科,吉林132011

出　　处：《郑州大学学报（医学版）》2006年第6期1058-1060,共3页Journal of Zhengzhou University(Medical Sciences)

基　　金：河南省及郑州大学"ODA"贷款资助项目

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、hsp60DNA,插入pPRO-EX-HTb质粒,IPTG诱导B21E.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。Aim： To get efficient expression vector of mitochondrial matrix protein 70 and 60. Methods：The fulllength mitochondrial heat shock protein 70 and 60 （Hsp70 and Hsp60） DNA was amplified from a eDNA 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 immuni;~ed rabbit was obtained, and Western-blot analysis was done after being purified. Results and Conclusion： The pPRO-EX-HTb expression vectors have been construtted successfully. And antibodies have been acquired, which have laid the foundation for further research on the function of mitochondrial matrix protein.

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 

分 类 号：Q51[生物学—生物化学] Q78