表达载体pEGFP-C3-PENK的构建以及在NIH3T3细胞中的表达

Objective: To construct recombinant plasmid of pEGFP-C3-PENK, and identify the position and time that the plasmid was expressed. Methods: Link PENK with FL eukaryotic expression vector to construct recombinant plasmid (pEGFP-C3-PENK), and transfect it into NIH3T3 cells with low concentration of naked plasmid and high concentration of naked plasmid. Mediated by Liposome 2000, transfection rate and the expression of enkephalin protein were observed by cell immunochemist and radioimmunity. Results: Recombinant plastnid pEGFP-C3-PENK was constructed successfully; the transfection rate of low concentration of naked plasmid was about 5 %; the transfection rate of high concentration of naked plasmid was about 8%; transfection rate by Liposome 2000 was about 20%; and green-fluorescence protein was expressed for 5 weeks. Conclusion: Recombinant plasmid pEGFP-C3-PENK was constructed successfully, which can express in NIH3T3 cells for 5 weeks.