Expression of nonstructural protein 3AB gene of Foot-and-mouth disease virus in Pichia pastoris

To express nonstructural protein 3AB of foot-and-mouth disease virus(FMDV) in Pichia pastoris expression system in order to differentiate the infected and vaccinated animals and for the further study of 3AB protein function,the gene 3AB was amplified from the recombinant plasmid of pGEM-T-Easy-3ABC by PCR and inserted into the Pichia pastoris expression vector pPICZαA.The recombinant expression plasmid was then linearized and transformed into Pichia pastoris GS115 by electroporation method. The expressed protein with the induction 0.5% methanol was analysed by SDS-PAGE and ELISA.It was demonstrated that the recombinant Pichia pastoris expression plasmid pPICZαA-3AB was successfully constructed and transformed into Pichia pastoris GS115. SDS-PAGE and ELISA analysis of the culture supernatants showed that the 3AB protein of 19 kD was expressed in Pichia pastoris. The expressed 3AB protein was reactive with the 3AB protein McAb against FMDV. It was concluded that the FMDV nonstructural protein 3AB was expressed in Pichia pastoris successfully. The expressed 3AB protein shared reaction with the 3AB protein McAb against FMDV.