Cortactin功能性结构域GST融合蛋白的表达和纯化  

Expression and purification of cortactin functional domain proteins fused with GST

作　　者：郝清亚[1] 袁岱岳[1] 邵伟斌[1] 毛勤生[2] 朱建伟[2] 

机构地区：[1]南通大学第二附属医院肝胆外科,南通226001 [2]南通大学附属医院普外科

出　　处：《南通大学学报（医学版）》2012年第6期455-457,共3页Journal of Nantong University(Medical sciences)

基　　金：国家自然科学基金资助项目(30771126;30772106);教育部回国留学人员基金资助项目(2008-890);江苏省自然科学基金资助项目(BK2006058)

摘　　要：目的:制备cortactin功能性结构域(NTA和ABR)GST融合蛋白,用于后续研究NTA和ABR多肽分子对肿瘤细胞的侵袭转移能力的影响。方法:通过从cortactin基因cDNA克隆中扩增获得cortactin功能性结构域(NTA和ABR片段),经双酶切和连接后将其分别插入载体pGEX-4T-2中获得重组质粒。将重组质粒转化入感受态菌,诱导表达,蛋白粗提后纯化。结果:PCR扩增后获得和预期一致的cortactin功能性结构域(NTA和ABR片段),通过测序证实该两片段正确插入表达载体。纯化的蛋白作SDS-PAGE电泳,证实蛋白的完整性和纯度良好。结论:成功地获得cortactin功能性结构域(NTA和ABR)GST融合蛋白。Objective： To prepare the cortactin functional domain proteins fused with GST,so as to make a preparation for the further study on the effect of cortactin functional domain polypeptides on tumor cells invasion and metastasis formation.Methods： The fragments of NTA and ABR were acquired from clone amplification of cortactin gene by polymerase chain reaction（PCR）,and then were inserted into the vector pGEX-4T-2,after double digestion and ligation with restriction endoncleases.The correctly sequenced plasmids were transfected into competent colon bacillus to express polypeptides which were crude extracted and purified.Results： The correct NTA and ABR fragments were obtained.It was confirmed that the NTA and ABR fragments were inserted into the expression vector correctly by sequencing respectively.The purity and integrity of the purified proteins were confirmed.Conclusion： The cortactin functional domain proteins fused with GST were successfully prepared.

关 键 词：CORTACTIN 重组质粒 蛋白 肿瘤 

分 类 号：R73-37[医药卫生—肿瘤]