军医进修学院学报2011，Vol.32Issue(1)：9-11,14,4.
大鼠FasL全长cDNA真核质粒的构建及转染HepG2细胞体外诱导细胞凋亡的作用
Construction of eukaryotic plasmids of rat FasL full length cDNA and role of FasL transfection in induction of in vitro apoptosis of HepG2 cells
摘要
Abstract
Objective To construct the eukaryotic expression vector and observe the effect of FasL gene transfection on induction of apoptosis of HepG2 cells. Methods RT-PCR and TA cloning technique were used to amplify the rFasL full length cDNA from rat testis cells, and then a eukaryotic expression vector pDC315 containing rFasL cDNA was constructed. Plasmid was transfected into HepG2 cells. The cells were harvested after 48h to identify the expression of plasmids by RT-PCR and Western blot. Meanwhile, two groups of HepG2 cells were respectively transfected with pDC315 plasmid as a plasmid control group and with H2O as a blank control group. After 48h of culture, the three groups of cells were counted, compared and analyzed. Results The sequence of cloned rFasL cDNA was consistent with that in GenBank. The HepG2 cells transfected with rFasL eukaryotic expression vector could express rFasL mRNA and protein. A large number of HepG2 cells died in the pDC315-rFasL experimental group. Conclusion The rFasL cDNA can be cloned and its eukaryotic expression vector can be constructed, indicating that rFasL gene can be expressed in HepG2 cells transfected with pDC315-rFasL. HepG2 cells expressing FasL induce apoptosis of neighbor-or auto-HepG2 cells, leading to their death.关键词
Fas配体蛋白质/克隆细胞/遗传载体Key words
Fas Ligand Protein/ Clone Cells/ Genetic Vectors分类
生物科学引用本文复制引用
许彪,胡瑾华,徐东平,李晓东,刘妍,陈婧,王业东,王慧芬..大鼠FasL全长cDNA真核质粒的构建及转染HepG2细胞体外诱导细胞凋亡的作用[J].军医进修学院学报,2011,32(1):9-11,14,4.基金项目
国家自然科学基金项目(30571671) （30571671）
军队"十一五"国际合作基金(06H057) （06H057）
