hTSHR膜外区氨基端基因原核表达载体的构建和鉴定  

Constructing and identifying the prokaryotic expressive vector of amino domain on the extracellular domain of human thyrotropin receptor cDNA

作　　者：陈慧[1] 方佩华[1] 陆凤先[1] 李宁[1] 吕枚[1] 许静[1] 

机构地区：[1]天津医科大学总医院核医学科,天津300052

出　　处：《天津医科大学学报》2005年第2期168-170,174,共4页Journal of Tianjin Medical University

基　　金：天津市科委自然科技基金攻关项目(编号:033605811)。

摘　　要：目的:为研究人促甲状腺素受体膜外区氨基端(hTSHR462bp)肽段的免疫学特性,将hTSHR462bpcDNA片段在原核表达系统中表达,故构建原核表达载体。方法:以人Graves病患者甲状腺组织中提取mRNA为模板,利用RT-PCR方法扩增双链甲状腺组织cDNA,又以PCR技术扩增目的基因,定向克隆法构建融合基因的原核表达载体pET102-hTSHR462bp。结果:获得了重组目的基因原核表达载体并经PCR鉴定、酶切鉴定插入片段方向、大小正确,DNA测序分析表明插入处接头和读框与预期序列相符。结论:成功构建了融合基因的原核表达载体pET102-hTSHR462bp,为进一步的蛋白表达、其生物学功能研究和未来临床应用奠定了基础。Objective To clone the amino terminal region of human thyrotropin receptor cDNA and construct its prokaryotic expressive vector. Methods hTSHR gene fragment had been cloned from thyroid tissues of Graves’disease patient by RT-PCR and inserted into pET102 vectors. After identificating by restriction analysis and sequencing it was confirmed that the interesting gene had been inserted into the vector in correct orientation and the right coding region. Results Afeter identifying by restriction analysis and seqencing it was confirmed that the interesting gene had been inserted into the vector in correct arientation and right coding region. Conclusion The plasmid pET 102-hTSHR 462bp will be useful in the epitope analysis of the region.

关 键 词：人促甲状腺素受体膜外区氨基端 基因克隆 测序 载体构建 

分 类 号：Q78[生物学—分子生物学]