中国药理学通报2016，Vol.32Issue(12)：1767-1771,1772,6.DOI:10.3969/j.issn.1001-1978.2016.12.025
pEGFP-NTCP稳定转染HEK293细胞系的建立及鉴定
Establishment and identification of HEK293 cell lines with stable and high expression of NTCP
摘要
Abstract
Abstrate:Aim To construct HEK293 cell line with stable and high expression of sodium taurocholate cotransporting polypeptide (NTCP ) efficiently and rapidly.Method Vector expressing EGFP-NTCP fusion protein was constructed and verified by DNA sequencing.The pEGFP-NTCP expression vector was transfected into HEK293 cells by FuGENE 6 transfection reagent. The transfected cells with high expression of green fluorescent protein were selected using fluorescence microscope for screening of G418 for 14 days to obtain cell lines stably and highly expressing NTCP.NTCP expression was detected by RT-PCR,qRT-PCR, Western-blot and the uptake experiment of taurocholic acid.Re-sult RT-PCR,qRT-PCR,Western-blot and the uptake experi-ment revealed that compared to the control cells,the expression of NTCP was significantly positive (P<0.01)in stable trans-fected cells showing green fluorescence (P<0.05 ).Conclusion The HEK293 cell line with stable and high expression of NTCP has been established efficiently and rapidly,which provides a cellular model for the study of the mechanism of the uptake of bile acid derivatives.关键词
NTCP/EGFP/稳定转染/牛磺胆酸/HEK293细胞/高效快速Key words
NTCP/EGFP/stable transfection/taurocholic acid/HEK293 cells/efficient and fast引用本文复制引用
陈雅,李静,张文政,罗敏,傅晓钟,刘亭..pEGFP-NTCP稳定转染HEK293细胞系的建立及鉴定[J].中国药理学通报,2016,32(12):1767-1771,1772,6.基金项目
国家自然科学基金资助项目（No 81460523）；贵州省优秀青年科技人才培养对象专项基金（No 2013-45号）；贵州省社会发展攻关计划项目（No 2013-3031号）；贵州省中药现代化科技产业研究开发专项（No 2013-4001号）；贵州省科学技术基金 （）
