拟南芥Ran2基因的原核表达及产物的纯化  

Expression in E.coli of Ran2 Gene from Arabidopsis thaliana and Purification of Expressed Products

作　　者：马立安[1,2] 江涛[3] 张忠明[4] 

机构地区：[1]长江大学生命科学学院 [2]华中农业大学农业微生物学国家重点实验室,湖北武汉430070 [3]长江大学动物科学学院 [4]华中农业大学农业微生物学国家重点实验室

出　　处：《长江大学学报（自科版）（中旬）》2007年第1期45-47,60,共4页Journal of Yangtze University(Nature Science Edition)

基　　金：国家自然科学基金项目(30070370)

摘　　要：采用RT-PCR方法扩增Ran2cDNA完整的编码区序列,构建pTYB2-Ran2重组质粒,转化大肠杆菌ER2566,经IPTG诱导蛋白表达后SDS-PAGE分析表达产物及存在形式。结果表明,表达的蛋白26ku与预期的理论值一致,并以包涵体形式存在;包涵体经纯化、破碎变性、复性及PBS透析,得到纯度较高的表达蛋白(Ran2)。The coding sequence of Ran2(666 bp)for protein expression was amplified by RT-PCR technique.Then the recombinant plasmid pTYB2-Ran2 was constructed.The recombinant protein was expressed in E.coli ER 2566 after induced by IPTG.SDS-PAGE was used to analyze expression and existent position of target protein.The results showed that there was a specific band at about 26 ku in size,which was identical with the expected molecular weight of target protein and in Pellet of inducing in ER2566.The more purified target proteins were obtained by purifying,denaturalizing,renaturing of Pellet.

关 键 词：拟南芥(Arnbidopsis thaliana) 小GTP结合蛋白 Ran2基因 原核表达 蛋白纯化 

分 类 号：Q784[生物学—分子生物学]