南方医科大学学报Issue(3)：308-311,4.DOI:10.3969/j.issn.1673-4254.2014.03.04
HERG基因G572R突变体表达载体的构建及其稳定转染细胞系的建立
Construction of pcDNA3-HERG-G572R expression vector and establishment of a cell line stably expressing HKE-HERG-G572R
摘要
Abstract
Objective To construct the pcDNA3-HERG-G572R expression vector and establish a cell line stably expressing HKE-HERG-G572R. Methods HERG-G572R mutant fragment was constructed by over-lap extension PCR and validated by DNA sequencing. The HKE-HERG-G572R expression vector was constructed and transfected into HEK293 cells to obtain a cell line stably expressing HKE-HERG-G572R. Results The pcDNA3-HERG-G572R expression vector was successfully constructed and the cell line stably expressing HKE-HERG-G572R was established. Real-time PCR and Western blotting revealed a 632-fold HKE-HERG-G572R overexpression in the transfected HEK293 cells as compared with that in control HEK293 cells transfected with pcDNA3 (P<0.01).Conclusion The protocol can be used to construct the cell line stably expressing HKE-HERG-G572R to provide a cell model for studying individualized therapy.关键词
HERG基因/HEK293/转染/突变体Key words
HERG gene/HEK293 cell/transfection/mutation引用本文复制引用
杨阳,黄娜,高岭,常素娥,郭波,胡丽丽,宋土生,黄辰..HERG基因G572R突变体表达载体的构建及其稳定转染细胞系的建立[J].南方医科大学学报,2014,(3):308-311,4.基金项目
国家自然科学基金中加合作项目(81161120549)；陕西省科技攻关项目重大科技专项（2010ZDKG-50）@@@@Supported by National Natural Science Foundation of China (81161120549) （81161120549）
