Abstract: Abstract: 【Objective】: Construct the eukaryotic expression vector of pcDNA5/ FRT-nanog-delta48 and detect its expression in liver cancer cells 7721.【Method】: Clone the alternatively-spliced variant of nanog (nanog-delta48 )full-length coding sequence by RT-PCR, then insert the right PCR product into pMD18-T vector, after DNA analysis, subclone the correct identification into pcDNA5/FRT. The eukaryotic expression vector of pcDNA5/FRT-nanog-delta48 was transiently transfected into human liver cancer cell 7721 by liposome mediated, then identify its expression by RT-PCR and assess its proliferation by MTT.【Result】:The full-length coding sequence of human nanog-delta48 was cloned succefully.The eukaryotic expression vector of pcDNA5/ FRT-nanog-delta48 was proved to be constructed successful by DNA analysis, and it overexpressed in 7721 cells. MTT result showed cell proliferation increased after transfection.【Conclusion】:The alternatively-spliced variant of nanog(nanog-delta48) has similar activity to that of the full length version. It will lay the foundation for the further study of its own functions and the relationship with nanog of full-length

Key words: Human, nanog gene, alternatively-spliced variant, gene cloning, eukaryotic expression, transient transfection