c-fos启动子与GFP重组质粒的构建及在pc12细胞中的表达

Expression of GFP gene under the control of c-fos promoter at the recombinant p lasmid in pc12 cell

目的:构建c-fos启动子与GFP的重组质粒载体,并使其在体外培养哺乳动物pc12细胞中表达.方法:XbaI、HindIII双酶切含c-fos启动子的HF443质粒,纯化后与同样双酶切的绿色荧光蛋白基因连接;利用LipofectAMINE2000将重组质粒转染体外培养哺乳动物pc12细胞.结果:加入Na+通道激活剂乌头碱后,转染后的pc12细胞可发出较强的绿色荧光.结论:c-fos启动子与GFP重组质粒构建成功,并可用于哺乳动物活细胞c-fos基因的检测.

Aim: To express GFP gene under c-fos promoter at the recombinant plas mid in pc12 cell. Methods: The plasmid HF443, was digested by Hi nd III and Xba I , and ligated with GFP gene. The recombinant plasmid was transfected into pc12 c ell by Lipofect AMINE2000. Aconitine, a sodium channel activator, was used to in duce the expression of GFP gene in recombinant plasmid. Results: Green fluoresce nce was detected in the transfected cell under the fluorescent microscope after aconitine induction. Conclusion: The recombinant plasmid of GFP gene under the c ontrol of c-fos promoter has been established as the vector for studing the GFP gene in living cell.