Cloning of VP2 Gene of Canine Parvovirus and Expression in Pichia pastoris

Objective To establish a yeast Pichia pastoris expression system expressing canine parvovirus VP2 protein(CPV VP2) to serve the diagnosis of CPV,development of CPV vaccine and research of VP2 protein function.Methods CPV VP2 gene was amplified by PCR,cloned into secretory Pichia pastoris expression vector pPICZαA,and was constructed into eukaryotic expression vector,which was named pPICZαA-VP2.After being linearized with enzyme digestion,the vector was transformed into Pichia pastoris GS115 by electroporation method.The CPV VP2 protein expressed with methanol induction.The expressed protein in yeast was analyzed by SDS-PAGE and Western blotting.Results CPV VP2 gene was successfully amplified,SDS-PAGE and Western blotting analysis of the culture supernatants showed that the VP2 protein of 64.35 kD was expressed in Pichia pastoris,and the expressed CPV VP2 protein shared reaction with positive serum against CPV.Conclusion CPV VP2 has been successfully expressed in yeast Pichia pastoris.The expressed CPV VP2 protein shares reaction with positive serum against CPV.