中国医科大学学报2012，Vol.41Issue(3)：204-207,4.
高GChTwist融合蛋白载体构建及其表达条件优化
Cloning of GC-rich hTwist Preparation, and Charactirization of Fusion Protein
摘要
Abstract
Objective To construct expression vector of fusion protein GST/TWIST with GC-enriched in Twist, induce expressly, and purify GST/TWIST fusion protein for further studies like GST Pulldown experiment. Methods The GC- rich Twist fragments were amplified by polymerase chain reaction (PCR)with a strategy of 'multiple factors-employed together' and cloned into the expression vector pGEX-5X-l. Recombinant vector of pGEX-5X-l-Twisi was identified by restriction enzyme digestion and sequencing. The pGEX-5X-l-Twist recombinant was transformed into E. Coli BL21 .induced by 1PTG. TWIST fusion protein was purified by glutathione beads. The purified product was analyzed by SDS-PAGE and Western blot. Results The GC-rich Twist recombinant pGEX-5X-l-Twist had been successfully constructed. A band corresponding to specific MW protein can be detected by SDS-PAGE after induction. After purification,fusion protein with high purity was harvested. Conclusion The full length gene fragments of human GC-rich Twist were successfully cloned into prokaryotic expression vector. The optimal conditions for inducing the target protein were confirmed and the GST/TWIST fusion protein with high purity was obtained. This study provided a basis for the further research on the biological function of TWIST protein.关键词
高GChTwist/克隆/融合蛋白/原核表达Key words
GC-rich hTwist/ cloning/ recombinant protein/ prokaryotic expression分类
生物科学引用本文复制引用
王利利,刘彩红,朱亚勤..高GChTwist融合蛋白载体构建及其表达条件优化[J].中国医科大学学报,2012,41(3):204-207,4.基金项目
辽宁省自然科学基金项目(20092123) （20092123）
辽宁省高校科研项目计划(2009S108) （2009S108）
