摘要
目的:建立稳定表达核仁素的H9C2大鼠心肌细胞株,为进一步研究核仁素在心肌细胞凋亡中的作用提供细胞模型。方法:将含有核仁素全长的质粒pCMV-Tag2B-Nuc用脂质体转染技术导入H9C2细胞,使核仁素在细胞内表达,用G418筛选出稳定表达核仁素的细胞株;用双酶切、DNA测序鉴定质粒,用RT-PCR及Western blot检测核仁素基因在靶细胞中的整合与表达。结果:经G418反复筛选的H9C2细胞株中,核仁素基因在mRNA及蛋白质水平表达均较野生型细胞及转空载体细胞升高。结论:用含核仁素全长的质粒转染H9C2大鼠心肌细胞,经筛选后可稳定表达核仁素。
Objective：To establish H9C2 cell line that can stably express the nucleolin,which can provide a cell line with which we can analyze the function of nucleolin in apoptosis of cardiomyocytes. Methods：The pCMV - Tag2B - Nuc plasmid was transfeeted into H9C2 cells with lipofectin,the stable transfectants screened by G418 and the expression of nucleolin was identified and analyzed by RT - PCR and Western blot. The pCMV - Tag2B - Nuc plasmid was confirmed by double - enzyme digestion and sequencing. Resuits：After G418 screening, compared with the control group, the mRNA and protein of nucleolin in transfected cell line were highly expressed, as demonstrated by the RT - PCR and Western blot analysis. Conclusion ： The rat H9C2 cell line stably expressing nucleolin was successfully constructed.
出处
《长治医学院学报》
2009年第2期88-90,共3页
Journal of Changzhi Medical College
基金
湖南省科技计划资助(2007JT3022)
