人内皮素受体A基因克隆及表达  

Clone and Expression of Human ET_A

作　　者：潘晓菲[1] 时丽丽[2] 谭初兵[2] 吕超君[1] 徐为人[2] 汤立达[2] 

机构地区：[1]天津医科大学基础医学院,天津300070 [2]天津药物研究院天津市新药设计与发现重点实验室,天津300193

出　　处：《生物技术通报》2014年第2期187-192,共6页Biotechnology Bulletin

基　　金：天津市应用基础及前沿技术研究计划(12JCYBJC18800);国家“重大新药创制”科技重大专项(2012ZX09105102,2011ZX-09401-009)

摘　　要：为了建立内皮素受体拮抗剂筛选系统,克隆人ET A基因,构建真核表达载体,并实现pTag-Lite SNAP-ET A在CHOK1细胞中的瞬时表达。从人肺腺癌细胞系A549中克隆人ET A基因,连接到pTag-Lite SNAP质粒,构建表达载体pTag-Lite SNAPET A,用FuGENER HD转染试剂将表达载体pTag-Lite SNAP-ET A转染入CHO-K1细胞内,通过荧光显微镜检测融合蛋白SNAP-ET A的表达。DNA测序结果表明pTag-Lite SNAP-ET A表达载体构建成功,荧光显微镜检测结果表明人ET A在CHO-K1细胞中有效表达。It was to facilitate inhibitor screening of endothelin receptor, cloning of human endothelin receptor gene ET A, construction of eukaryotic expression vectors and transient expression of pTag-Lite SNAP-ET A in CHO-K1 cells. Method Human ET A gene fragment were amplified from human lung cancer cell line A549 via RT-PCR and inserted into pTag-Lite SNAP plasmid, the recombinant plasmids were then transiently transfected into CHO-K1 cells using FuGENE？ HD, expression of ET A was observed via fluorescence microscope. Result Sequencing result showed pTag-Lite SNAP-ET A expression vector were correctly constructed. Fluorescence microscope images indicated that two vectors were expressed in CHO-K1 cells.

关 键 词：内皮素受体A RT-PCR pTag-Lite SNAP载体 CHO-K1细胞 表达 

分 类 号：R346[医药卫生—基础医学]