Construction and Expression Detection of Eukaryotic Expression Vector of Green Fluorescent Protein Gene

We constructed the eukaryotic expression vector pcDNA3GFP for determination of expression of green fluorescent protein (GFP) .GFP cDNA was obtained from plasmid pCIGFP with single Not Ⅰ digestion and was inserted into the same restriction site of pcDNA3GFP. The upright insertion was determined with BamH Ⅰ digestion. With transfection of pcDNA3GFP encapsulated by LipofectAMINTM in- to the cultured MDCK cells, G418 was used for selection of positive clones exprssing green fluorescence. It was demonstrated that GFP was sucessfully expressed in MDCK cells, implying that GFT, was a good reporter gene and selection marker molecule.