医学分子生物学杂志2012，Vol.9Issue(3)：189-193,5.DOI:10.3870/j.issn.1672-8009.2012.03.007
小鼠GITRL基因真核表达质粒的构建转染及其在Kupffer细胞中的表达
Construction and Transfection of Eukaryotic Expression Plasmid Containing mGITRL Gene and Its Expression in Kupffer Cells
摘要
Abstract
Objective To construct eukaryotic expression plasmid pEGFG-Nl-GITRL containing mouse GITRL gene and transfer to the Kupffer cells in mice in vitro. Methods GITRL gene was amplified by PCR, and then cloned into pEGFP-Nl vector. The positive clones were selected for sequencing. Finally the GITRL gene was transfected into Kupffer cells by liposome -chemical method. Results The eukaryotic expression plasmid pEGFP -Nl -GITRL was constructed successfully , and its sequences were consistent with the published once in Genbank . After transient transfection of pEGFP-Nl -GITRL into the Kupffer cells in vitro, RT-PCR and WB revealed the stable expression of GITRL in Kupffer cells. Conclusion The eukaryotic expression plasmid pEGFP-Nl -GITRL was successfully constructed and expressed in mouse Kupffer cells , which may provide the foundation for further study on the biological function of GITRL in Kupffer cells.关键词
糖皮质激素诱导的肿瘤坏死因子配体/pEGFP-N1/Kupffer细胞/重组质粒Key words
glucocorticoid-induced tumor necrosis factor receptor ligand/pEGFP-N1/Kupffer cells/recombinant plasmid分类
医药卫生引用本文复制引用
张艳,李星,吴剑,罗静,龚建平..小鼠GITRL基因真核表达质粒的构建转染及其在Kupffer细胞中的表达[J].医学分子生物学杂志,2012,9(3):189-193,5.基金项目
国家自然科学基金(No.307702098,801070374) （No.307702098,801070374）
