人TSHR膜外区cDNA克隆及真核表达载体的构建  

cDNA cloning and construction of human TSHR ectodominant eukaryotic expression vector

作　　者：武敏[1] 施秉银[1] 伍丽萍[1] 兰玲[1] 吴小燕[1] 张进安[1] 徐莉[1] 

机构地区：[1]西安交通大学医学院第一附属医院内分泌科,陕西西安710061

出　　处：《西安交通大学学报（医学版）》2007年第2期123-125,共3页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.30371335);陕西省科技攻关项目(No.2002K10-G5)

摘　　要：目的通过生物技术获得促甲状腺激素受体膜外区表达载体,为TSHR膜外区的研究提供有力工具。方法从人甲状腺组织中提取总RNA,RT-PCR分离得到TSHR膜外区cDNA,双酶切后插入真核表达载体pcDNA3.1(+)。结果经酶切鉴定、测序分析表明TSHR膜外区片段与GenBank提供的完全相同。结论成功构建了人TSHR膜外区真核表达载体。Objective To construct the thyrotropin receptor （TSHR） eukaryotic expression vector by biological technique, which lays the foundation for further study of TSHR structure and function. Methods Total RNA was extracted from human thyroid, ETSHR cDNA was obtained with RT-PCR technique and subcloned into the plasmid pcDNA3. 1 （ ＋ ）. Results Evidence of DNA sequence analysis and restriction enzymes digestion showed that the fragment was the same as the one provided by GenBank. Conclusion THSR ectodominant gene was cloned into the eukaryotic expression vector pcDNA3.1 （＋）.

关 键 词：人TSHR膜外区 RT—PCR 真核表达载体 

分 类 号：R581[医药卫生—内分泌]