人TAT-PDX-1融合蛋白在大肠杆菌中的表达与纯化  

Expression and purification of human TAT-hPDX-1 fusion protein in E.coli

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

机构地区：[1]郑州大学第一附属医院,郑州450052

出　　处：《山东医药》2011年第19期18-20,共3页Shandong Medical Journal

基　　金：河南省杰出青年科学基金资助项目(074100510001)

摘　　要：目的构建人TAT-PDX-1表达载体,并对其融合蛋白进行纯化。方法采用RT-PCR技术从人胰腺组织中获得目的基因hPDX-1,将TAT序列与PDX-1克隆到原核表达载体pET-28a;用IPTG诱导、表达融合蛋白;NiNTAagaros纯化融合蛋白,Western blot鉴定。结果目的片段PDX-1被有效扩增,DNA序列测定表明所构建重组质粒pET28a-TAT-h PDX-1与设计相同;TAT-PDX-1融合蛋白在大肠杆菌中获得高效表达并成功纯化。结论成功地获得了TAT-hPDX-1融合基因的表达产物,为进一步研究及临床应用奠定了基础。Objective To construct a combinated vector containing TAT and human pancreatic duodenal homeobox-1（hPDX-1）,and to purify its fusion protein.Methods The target gene hPDX-1 was obtained from human pancreatic tissue by RT-PCR,and then PDX-1 and TAT were cloned to expressive vector pET-28a by;fusion protein was induced and expressed by IPTG;fusion protein was purified by Ni-NTA agaros,and detected by western blot.Results PDX-1 was effectively amplified and the TAT-PDX-1 gene sequencing showed sequence as scheduled;the fusion protein was successfully expressed in E.coli and purified.Conclusions Expression product of TAT-hPDX-1 fusion gene can be obtained successfully,and the results may lay a good basis for manufacture and clinical application of the TAT-hPDX-1.

关 键 词：原核表达 蛋白转导域 胰十二指肠同源盒基因-1 

分 类 号：R743[医药卫生—神经病学与精神病学]