Construction of Fluorescence Expression Vector of Ubiquitin-fold Modifier Conjugating Enzyme(UFC1) Gene

To construct eukaryotic expression plasmid of ubiquitin-fold modifier conjugating enzyme(UFC1) gene linked with GFP gene with pEGFP-C3 vector and observe its expression and location in eukaryotic cells,thus it can provide a powerful tool to further study the function of UFC1 gene.The full-length cDNA of UFC1 gene was amplified from human cervical carcinoma Hela cell by RT-PCR;UFC1 gene was cloned into eukaryotic expression vector pEGFP-C3 after double-digestion;To construct and identify the recombinant plasmid pEGFP-C3-UFC1.Then the plasmid was transfected into Hela cells by liposome,and its expression was detected by observing the expression of enhanced green fluorescent protein(EGFP)through fluorescent microscope.Recombinant pEGFP-C3-UFC1 vector was confirmed by double-digestion and sequencing.The green fluorescence was observed in transfected Hela cells.It was mostly located in the cytoplasm and distributed unequally with pEGFP-C3-UFC1 vector transfected;but it was located in cell whole and distributed equally with control vector pEGFP-C3 transfected.UFC1 gene was successfully cloned and recombinant pEGFP-C3-UFC1 vector was successfully constructed.