Construction and determination of a recombinant eukaryotic plasmid pcDNA3.1(+)-rmCGβ

Objective To construct an eukaryotic expression plasmid expressing macaca mulatta chorionic gonadotropin hormone β subunit(rmCGβ) gene and test whether the gene could be expressed in the cell B16 in vitro.Methods The full-length cDNA of rmCGβ gene was obtained by PCR.Then the cDNA was inserted into the eukaryotic expression vector pcDNA3.1(+).The recombinant eukaryotic plasmid pcDNA3.1(+)-rmCGβ was transfected into eukaryotic cell B16 by Lipofectin.RT-PCR was used to certify whether the gene was correctly expressed in B16/ cDNA3.1(+)-rmCGβ cell.Results cDNA of rmCGβ was correctly amplified.The recombinant eukaryotic expression plasmid pcDNA3.1(+)-rmCGβ was successfully constructed,and it could be correctly expressed in the B16 cells.Conclusion This recombinant plasmid pcDNA3.1(+)-rmCGβ will provide a basis for further study on unknown functions of rmCGβ.