Construction and Expression of Eukaryotic Expression Vector Bearing Fusion Gene of Human LIF Gene and EGFP

Objective To construct a recombinant eukaryotic vector bearing the fusion gene of human LIF gene and EGFP(enhanced green fluorescent protein),and to transfect it into human embryonic lung fibroblasts(hELF).Methods The LIF gene was amplified by PCR with the recombinant plasmid pcDNA3.1/ lif as the template and the fragment was combined with plasmid pEGFPN1.The recomninant expression vector,pEGFP-Nl/lif,was transfected to hELF using Lipofectamin.The expressed fusion protein was examined by fluorescence microscopy,RT-PCR and Western-blot.Results The eukaryotic expression vector pEGFP-N1/lif was successfully constructed.The fusion protein EGFP-LIF was detected in hELF.Conclusion The successful construction and expression of the eukaryotic expression vector pEGFP-N1/lif has been achieved.