中南民族大学学报（自然科学版）Issue(2)：36-40,5.
海南斑等蝎钾通道阻断剂 Im58的克隆、表达、纯化和鉴定
Cloning, Expression, Purification and Identification of Im58, the Potassium Channel Blocker of Scorpion Isometrus maculates
摘要
Abstract
To express potassium channel blocker Im58, full length DNA sequences of Im58 was amplified by overlap PCR with four designed primers and the cloning with the right sequence was transformed to E. coli/Rosetta ( DE3 ) by vector pGEX-4T-1. Under different inducing time and IPTG concentrations, the expressing quantities of fusion protein were investigated to optimize the inducing conditions. Then the recombinant GSH-fusion proteins were purified by GSH affinity chromatography, treated with enterokinase and purified by C18 column of RP-HPLC. The sample of Im58 purifying peptides was obtained to identify the molecular weight and purity by SDS-PAGE Gel of Tricine system. Accurate molecular weight was also identified by MALDI-TOF-MS. The results showed that the prokaryotic expression plasmid of Im58 was successfully constructed by positive clone. Moreover, the expression, purification and identification of Im58 were confirmed by SDS-PAGE Gel of Tricine system and MALDI-TOF-MS.关键词
钾离子通道阻断剂/Im58/原核表达/蛋白纯化Key words
potassium channel blocker/Im58/prokaryotic expression/protein purification分类
化学化工引用本文复制引用
尹世金,兰珍,李羽欣..海南斑等蝎钾通道阻断剂 Im58的克隆、表达、纯化和鉴定[J].中南民族大学学报（自然科学版）,2013,(2):36-40,5.基金项目
国家自然科学基金资助项目(30900239) （30900239）
广东省产学研资助项目(2010B090400047) （2010B090400047）
