Expression of VP2 Gene of Feline Panleukopenia Virus Isolated from Tiger in Pichia pastoris Yeast / 微生物学通报

ABSTRACT

In this study,the VP2 gene of feline panleukopenia virus(FPV) isolated from tiger was successfully cloned and expressed in Pichia pastoris yeast.The VP2 gene was amplified by PCR with a pair of specific primer.Then PCR products was purified and cloned into pGEM-T for sequencing.The interesting gene fragment was recovered after the double enzyme digestion of EcoRI/NotI,then subcloned into pPICZ?A for secretory expression.The recombinant pPICZ?AVP2 was linearized with SacI and then transformed into competence yeast GS115 for expression under the induction of 1% methanol.The positive recombinants were screened by PCR method.The expression product was identified by SDS-PAGE and western-blotting.The results showed that there was a molecular weight of 32 kD,which could be specifically recognized by polyclonic antibody against FPV.It revealed that the recombinant VP2 protein had correct three-dimensional structure,which would be used as a antigen protein for the diagnosis and prevention of FPV infection in tigers.