基础医学与临床Issue(2)：218-223,6.
RACK1过表达和低表达HUVEC细胞系的建立和鉴定
Establishment and identification of HUVEC cell strains with over-expression and low expression of RACK 1
摘要
Abstract
Objective To establish several human umbilical vein endothelial cell ( HUVEC ) strains with over-ex-pression or low expression of receptor for activated C kinase 1 ( RACK1 ) , which will provide an effective tool for future studying the function of RACK1 in arrhythmia.Methods The full-length cDNA sequence of RACK1 gene was amplified and inserted into pIRES2-EGFP.At the same time, designed and synthesised complementary DNA sequences of 3 pairs of short hairpin structure and a pair of negative control sequence , then subcloned into the plas-mid pGenesil-1 .The HUVEC cells were transfected with these plasmids and screened by using G 418 .And the expression of RACK1 mRNA and protein in the cells were assayed by qRT-PCR and Western blot , respectively . Results RACK1 eukaryotic expression vector and siRNA expression vectors of RACK 1 were constructed success-fully.After a 48 h transfection of HUVEC cells with the recombinant vectors and G 418 selection, the positive cell clones were obtained .qRT-PCR and Western blot showed that over-expression vector and interference vectors could effectively enhanced and knocked-down RACK1 expression in HUVEC strains .Conclusions HUVEC cell strains with over-expression and low expression of RACK 1 have been successfully established .关键词
RACK1/过表达/RNA干扰/HUVEC细胞系Key words
RACK1/over-expression/RNA interference/HUVEC strain分类
生物科学引用本文复制引用
张丽,贾雄飞,牛华,冯悦,宋玉竹,张望龙,李胜营,吴明江,毛小琴..RACK1过表达和低表达HUVEC细胞系的建立和鉴定[J].基础医学与临床,2015,(2):218-223,6.基金项目
国家自然科学基金（81260248）；云南省应用基础研究计划 （）
