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.
出处
《生物技术通报》
CAS
CSCD
北大核心
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)
