Microbiology 1992;0(01):-

Expression of VP2 Gene of Feline Panleukopenia Virus Isolated from Tiger in Pichia pastoris Yeast

Songtao YANG

Keywords

Feline panleukopenia virus,VP2 protein gene,Pichia pastoris,Expression

Country

China

Language

Chinese

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.