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.
出处
《长江大学学报（自科版）（中旬）》
CAS
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)
