豌豆胰岛素基因克隆、表达及融合蛋白纯化  

Cloning of PA1b gene and its expression and purification of fusion protein

作　　者：杜雯[1] 徐平勇[1] 陈正望[1] 

机构地区：[1]华中科技大学生命科学与技术学院,湖北武汉430074

出　　处：《华中科技大学学报（自然科学版）》2004年第9期102-104,共3页Journal of Huazhong University of Science and Technology(Natural Science Edition)

基　　金：国家高技术研究发展计划资助项目 (2 0 0 2AA2 1 4 0 6 1 )

摘　　要：将人工合成的豌豆胰岛素 (PA1b)基因寡核苷酸片段 ,通过PCR扩增得到PA1b基因 ,将此基因克隆到原核表达载体 pMAL p2x中 ,然后将重组质粒pMAl p2X PA1b转化大肠杆菌DH5α ,经IPTG诱导进行可溶性表达 .以AmyloseResin亲和层析 ,纯化出融合蛋白MBP PA1b .经SDS PAGE及Western印迹证明 ,融合蛋白MBP PA1b在大肠杆菌中表达成功 。The gene fragment of PA1b was synthesized and cloned into prokaryotic expression vector pMAL p2x.The recombinant expression plasmid pMAL p2 PA1b was transformed into E.coli DH5α. By IPTG induction, the fusion protein MBP PA1b was produced and purified by Amylose Resin affinity column. The analysis of SDS PAGE and Western blot confirmed that the MBP PA1b fusion protein was expressed in E. coli, and it was useful for further investigation of PA1b.

关 键 词：豌豆胰岛素 表达 融合蛋白 克隆 

分 类 号：Q516[生物学—生物化学] Q786