Cloning and eukaryotic expression of mouse Uncv gene

Objective To clone the full length cDNA of Uncv gene in mice and to express the gene in eukaryotic cells.Methods RT-PCR assay was applied to clone the full length coding region of the Uncv gene and constructed its expression plasmid pcDNA 3.1-Flag/Uncv.The recombinant plasmid was transfected into HeLa cells and the fusion protein was identified by Western blot analysis.Results The complete coding sequence was obtained and cloned into the pcDNA 3.1-Flag vector.The recombinant pcDNA 3.1-Flag/Uncv plasmid was transiently expressed in HeLa cells.HeLa cell clones expressing fusion protein with molecular weight of about 95×103 were obtained.Conclusions A recombinant eukaryotic expression plasmid of Uncv has been successfully constructed and expressed in HeLa cells.It may provide a foundation for further biological studies of Uncv gene.