PMEPA1蛋白的原核表达、纯化及其抗体制备*

J4 ›› 2012, Vol. 47 ›› Issue (3): 10-13.

PMEPA1蛋白的原核表达、纯化及其抗体制备*

许丽红,罗深恒,王莎莎,侯国丽,刁爱坡

天津科技大学生物工程学院, 天津 300457

收稿日期:2011-12-30 出版日期:2012-03-20 发布日期:2012-04-01
通讯作者: 刁爱坡(1971- ),男,博士,教授,研究方向为细胞生物学. Email:diaoaipo@tust.edu.cn
作者简介:许丽红(1986- ),女,硕士研究生,研究方向为细胞分子生物学. Email:xlh267026947@126.com
基金资助:
天津市自然科学基金重点项目(10JCZDJC16800)

Expression and purification of PMEPA1 protein and its antibody preparation

XU Li-hong, LUO Shen-heng, WANG Sha-sha, HOU Guo-li, DIAO Ai-po*

School of Biological Engineering, Tianjin University of Science and Technology, Tianjin 300457, China

Received:2011-12-30 Online:2012-03-20 Published:2012-04-01

摘要/Abstract

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。

关键词: GST-PMEPA1; His-PMEPA1; 原核表达; 抗体; 纯化

Abstract:

To study expression and function of PMEPA1 protein in cells, the constructs of pTrcHisA-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 respectively. A rabbit was immunized with purified His-PMEPA1, the antibody against PMEPA1 was prepared successfully, and purified using GST-PMEPA1. Western blotting showed that the purified antibody can recognize endogenous PMEPA1 proteins in cell lines.

Key words: GST-PMEPA1; His-PMEPA1; prokaryotic expression;antibody; purification

许丽红,罗深恒,王莎莎,侯国丽,刁爱坡. PMEPA1蛋白的原核表达、纯化及其抗体制备*[J]. J4, 2012, 47(3): 10-13.

XU Li-hong, LUO Shen-heng, WANG Sha-sha, HOU Guo-li, DIAO Ai-po*. Expression and purification of PMEPA1 protein and its antibody preparation[J]. J4, 2012, 47(3): 10-13.

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