Expression of 3AB protein of foot and mouth disease virus in Pichia pastoris

Abstract

3AB non structural protein (nsp) was used to diagnose the vaccinated animals from those infected with foot and mouth disease virus (FMDV). In order to express the gene encoding 3AB protein of FMDV type A22 in Pichia pestoris, the gene was amplified and cloned into the yeast transfer vector (pPIC-9K) at EcoRI site. The cloned gene was further characterized by colony PCR, restriction enzyme digestion and sequence analysis. The recombinant plasmid was transferred into GS115 strain of P. pastoris cells by electroporation. The His⁺ Pichia transformants were analyzed for the presence of the insert in the yeast genome by PCR. PCR positive clones were grown and expression was induced with 0.5% methanol. The expressed gene products were then characterized by SDS-PAGE and Western blot analysis. This is the first report on the production of FMDV non structural proteins in yeast. The expressed protein will be of diagnostic importance.