Construction of eukaryotic expression vector of bombyxinII

Objective To construct an eukaryotic expression vector of bombyxinⅡin order to provide a basis for further study of the hypoglycemic effects in mammalian cell.Methods BbxⅡ cDNA was amplified by PCR.Plasmid PUC 57 which contain BbxⅡ cDNA was used as template.The fragment was digested and ligated to the eukaryotic expression vector PcDNA3.1 The ligation mixture was transformed into competent E.coli DH-5α cells.Transformants containing inserts were confirmed by restrictive digestion and DNA sequencing.Results The PCR product was about 300 bp.The recombinant expression vector was identified by restrictive digestion and DNA sequencing.Conclusion The recombinantⅡ eukaryotic expression vector PcDNA3.1-BbxⅡ/His is constructed successfully.