Construction of human TIMP-2 eukaryotic expression plasmid and its expression in vitro

Objective To construct a eukaryotic expression vector of human TIMP-2 gene and to gain a stable expression in vitro by transfecting it into CHO cell.Methods The recombinant eukaryotic expression plasmid pcDNA3/TIMP-2 was constructed by ligating human TIMP2 gene,which was amplified by RT-PCR and the pcDNA3 vector.The recombinant plasmid was transfected into CHO cells by liposome,the expression of TIMP-2 was verified by Western blot.Results The recombinant pcDNA3/TIMP2 plasmid was correctly constructed and confirmed by restriction endonuclease analysis and DNA sequencing.Western blot showed the expression of TIMP-2 protein.Conclusion The recombinant eukaryotic expression plasmid of TIMP-2 is successfully constructed,and a stable expressing cell line of it is established.