文章摘要
蒋玲琳,刘丽,朱含章,杨青,范乐明,陈琪.稳定表达GlyRα1的HEK293细胞系的建立[J].南京医科大学学报,2007,(7):647~650
稳定表达GlyRα1的HEK293细胞系的建立
Establishment of a stable GlyRα1-expressed HEK293 cell line
投稿时间:2007-01-08  
DOI:10.7655
中文关键词: 甘氨酸受体  基因转染  HEK293细胞  稳定表达
英文关键词: glycine receptor  gene transfection  HEK 293 cell  stable expression
基金项目:江苏省高校自然科学重大基础研究项目(06KJA31027)
作者单位
蒋玲琳 南京医科大学动脉硬化研究中心,江苏 南京 210029 
刘丽  
朱含章  
杨青  
范乐明  
陈琪  
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中文摘要:
      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?
英文摘要:
      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.
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