Construction of eukaryotic expression plasmid of APM1 gene and its expression In HEK 293 cells / 中华内分泌代谢杂志

ABSTRACT

Objective To construct an eukaryotic expression vector with human adipose most abundant gene transcript 1 (APM1) gene,and to investigate the transfection and expression of pCDEF-APM1 eukaryotic expression plasmid in HEK293 cells.Methods pCDEF-APM1 eukaryotic expression plasmid was constructed by DNA recombinant method.Expression vector pCDEF-APM1 was transfected into HEK293 cells with Effectene reagent.The level of human adiponectin protein in the supernatant of cell culture media was detected with double antibody sandwich ELISA.Results The sequence of DNA fragment from constructed pCDEF-APM1 plasmid was identical to that published in GenBank.There was raised human adiponectin protein level in culture supernatant of HEK293 cells tnmsfected with pCDEF-APM1.Conclusion The pCDEF-APM1,an eukaryotic expression plasmid for APM1 gene is successfully constructed.High protein expression of adiponectin can be obtained in HEK293 cells transfected with pCDEF-APM1 eukaryotic expression plasmid.