TAT-SOD融合蛋白表达载体的构建  

Construction of TAT-SOD fusion protein expression vector

作　　者：王茜茜[1] 宋波[2] 苗成[2] 韩志强[2] 许予明[2] 

机构地区：[1]郑州市第三人民医院神经内科,郑州450002 [2]郑州大学第一附属医院神经内科,郑州450052

出　　处：《郑州大学学报（医学版）》2010年第1期72-74,共3页Journal of Zhengzhou University(Medical Sciences)

基　　金：河南省杰出青年科学基金资助项目074100510001

摘　　要：目的:构建pET28a-TAT-SOD原核表达载体,纯化并测定其活性。方法:PCR法扩增TAT-SOD片段,插入载体pET28a后得到pET28a-TAT-SOD重组表达子,转化大肠杆菌BL21(DE3),IPTG诱导TAT-SOD融合蛋白表达。表达产物用SDS-PAGE电泳鉴定,组氨酸亲和层析柱纯化融合蛋白,利用SOD活性测定试剂盒测定其活性。结果:构建了高表达重组子pET28a-TAT-SOD,获得了相对分子质量约为25000的具有活性的融合蛋白TAT-SOD,纯度达90%。纯化的蛋白浓度为700mg/L,活性为197.54U/L。结论:成功构建了具有活性TAT-SOD融合蛋白的表达载体。Aim：To construct a vector of pET28-TAT-SOD,purify and detect its activity.Methods：TAT-SOD fragment was amplified,which then was inserted into pET28a vector to construct the expression vector of pET28a-TAT-SOD.The recombinant vector was transformed into E.coli BL21 and induced with IPTG.The highly expressed TAT-SOD was purified by affinity chromatography and the enzymatical activity was detected by SOD activity kit.Results：pET28a-TAT-SOD was successfully constructed,and fusion protein with relative molecular mass about 25 000 was expressed in prokaryotic cells.The purity of fusion protein was 90%,the concetration was 700 mg/L,and the activity was 197.54 U/L.Conclusion：pET28a-TAT-SOD has been successfully expressed in prokaryotic cells and fusion protein TAT-SOD has been successfully purified.

关 键 词：TAT SOD 蛋白转导 载体 

分 类 号：R730[医药卫生—肿瘤]