Expression of Fusion Protein of Goose Paramyxovirus in Pichia pastoris

To express F protein of NA-1 in Pichia pastoris,F gene was amplified by RT-PCR from NA-1 with a pair of specific primers.Then PCR product was purified and cloned into pGEM-T vector to obtain the plasmid pT-F.The gene fragment was recovered after the double enzyme digestion with EcoRⅠ and NotⅠ,then was subcloned into pPICZα A.The recombinant pPICZαA-F was linearized with PmeⅠ and then transformed into GS115 yeast cells for expression.The recombinant strains were screened by PCR technique.The expression products were identified by SDS-PAGE and Western blot.The results showed that there was a molecular weight of 63 ku protein specifically recognized by polyclonal antibody against NA-1.It suggested that the F protein of NA-1 was obtained in Pichia pastoris expression system.