白血病相关蛋白LRP16过表达HepG2稳定细胞系的构建及鉴定  

Construction and Identification of HepG2 Cell Line Stably Overexpressing Leukemia Related Protein 16

作　　者：李婷[1] 王安平[1] 李萍[1] 王双双[1] 母义明[1] 

机构地区：[1]解放军总医院内分泌科,北京100853

出　　处：《生物技术通讯》2015年第5期611-614,共4页Letters in Biotechnology

基　　金：国家自然科学基金(81471026)

摘　　要：目的:构建白血病相关蛋白16(LRP16)过表达的Hep G2稳定细胞系,鉴定LRP16过表达的效果。方法:把EX-Y2069-Lv201过表达质粒载体包装成LPP-Y2069-Lv201-400过表达慢病毒,感染Hep G2细胞系,通过荧光筛选及药物筛选,获得LRP16稳定过表达细胞系;应用q PCR和Western印迹鉴定该稳定细胞系中LRP16的表达。结果:q PCR结果显示过表达稳转株(LPP-Y2069-Lv201-400)中LRP16基因的表达量为对照稳转株(LPP-NEG-Lv201-100)的128.64倍;Western印迹结果显示过表达稳转株的LRP16表达量明显高于对照稳转株,结果与q PCR一致。结论:构建并鉴定了人LRP16基因过表达Hep G2稳定细胞系。Objective： To establish a HepG2 cell line over-expressing leukemia related protein 16（LRPI6） medi- ated by a lentiviral system and verify the stable over-expression profile of LRP16. Methods： The LRP16 over-ex- pression plasmid vector EX-Y2069-Lv201 was packaged to recombinant lentivirus LPP-Y2069-Lv201-400 and transfected HepG2 cells. Fluorescence detection and drug selection were used to screen transducted HepG2 cells. Real time qPCR and Western blotting were used to examine the over-expression profile of LRP16. Results： The qPCR result showed that the gene expression of LRP16 in constructed HepG2 cell line was 128.64-fold of the control cell. Western blotting results showed that the protein expression level of LRP16 in resulting HepG2 cell line was significantly higher than the control cell, which was consistent with qPCR. Conclusion： The HepG2 cell line stably overexpressed LRP16 was established with lentiviral system.

关 键 词：白血病相关蛋白LRP16 慢病毒载体 HEP G2细胞 

分 类 号：Q78[生物学—分子生物学]