稳定表达GlyRα1的HEK293细胞系的建立  

Establishment of a stable GlyRα1-expressed HEK293 cell line

作　　者：蒋玲琳[1] 刘丽[1] 朱含章[1] 杨青[1] 范乐明[1] 陈琪[1] 

机构地区：[1]南京医科大学动脉硬化研究中心,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2007年第7期647-650,共4页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省高校自然科学重大基础研究项目(06KJA31027)

摘　　要：目的:建立稳定表达甘氨酸受体α1亚基(GlyRα1)真核表达载体的人胚肾细胞(HEK293)细胞系。方法:构建pcDNA3.1-GlyRα1真核表达载体,应用脂质体介导的转染技术将该质粒导入HEK293细胞,再用G418筛选表达稳定的细胞系。真核细胞中GlyRα1的表达分别用RT-PCR与Western blot方法检测。结果:在7株转染并经G418反复筛选的HEK293细胞系中,有4株细胞系明显表达GlyRα1的mRNA及其蛋白质,其余3株细胞表达较弱或者没有表达。结论:用pcDNA3.1-GlyRα1转染的HEK293细胞经G418筛选,可成功建立GlyRα1稳定表达系。Objective：To establish a stable glycine receptor alpha 1 （GlyRα1）-expressed HEK293 cell line. Methods：The pcDNA3.1- GlyRα1 plasmid was constructed and transfected into HEK293 cells with lipofectin. The stable transfectants were screened by G418. The mRNA expression of GlyRα1 was identified using RT-PCR. The protein expression was analyzed by Western blot. Results： Selected by G418, 4 out of 7 transfected cell lines showed high expression level of GlyRα1, as demonstrated by RT-PCR and Western blot analysis. No or low expression of GlyRα1 was shown in other 3 cell lines. Conclusion：A HEK293 cell line stably expressing GlyRα1 was constructed successfully.

关 键 词：甘氨酸受体 基因转染 HEK293细胞 稳定表达 

分 类 号：Q813[生物学—生物工程]