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PMEPA1蛋白的原核表达、纯化及其抗体制备

Expression and purification of PMEPA1 protein and its antibody preparation
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摘要 为了从蛋白水平上检测前列腺跨膜蛋白PMEPA1(prostate transmembrane protain,androgen induced 1)在细胞内的表达和功能,构建了原核表达重组质粒pTrcHisA-PMEPA1和pGEX4T-2-PMEPA1,转化大肠杆菌BL21-RIPL后,IPTG诱导蛋白表达,分别用Ni-NTA和谷胱甘肽琼脂糖凝胶层析柱纯化蛋白。用His-PMEPA1免疫兔子制备抗体,并用GST-PMEPA1对抗体进行纯化。免疫印迹检测显示抗体能够特异识别细胞中内源PMEPA1蛋白。 To study expression and function of PMEPA1 protein in cells, the constructs of pTrcHlsA-PMEPA1 and pGEX4T-2-PMEPA1 were made and transformed into E. coli BL21-RIPL to express recombinant protein His-PMEPA1 and GSTpPMEPA1. The proteins were purified by Ni-TNA and Glutathione Sepharose 4B affinity chromatography re- spectively. A rabbit was immunized with purified His-PMEPA1, the antibody against PMEPA1 was prepared success- fully, and purified using GST-PMEPA1. Western blotting showed that the purified antibody can recognize endogenous PMEPAI proteins in cell lines.
作者 许丽红 罗深恒 王莎莎 侯国丽 刁爱坡
机构地区 天津科技大学生物工程学院
出处 《山东大学学报（理学版）》 CAS CSCD 北大核心 2012年第3期10-13,共4页 Journal of Shandong University(Natural Science)
基金 天津市自然科学基金重点项目(10JCZDJC16800)
关键词 GST-PMEPAl His-PMEPAl 原核表达 抗体 纯化 GST-PMEPA1 His-PMEPA1 prokaryotic expression antibody purification
分类号 Q78 [生物学—分子生物学]
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参考文献12

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二级参考文献25

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山东大学学报（理学版）
2012年 第3期

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