Expression of Capsid Precursor Polypeptide (PI) of Foot and Mouth Disease Virus Vaccine Strains in <i>Pichia pastoris</i>

Abstract

Foot and mouth disease virus (FMDV) is the aetiological agent of a highly contagious vesicular disease of c1ovenhooved animals. The gene coding for the capsid polyprotein (P1) of FMDV from vaccine strains was amplified and cloned into the Yeast transfer vector (pPIC-9K) at <i>Eco RI </i>and <i>Not I. </i>The cloned PI gene was characterized by colony PCR, restriction enzyme digestion and sequence analysis. The recombinant plasmid of each serotype was transferred

into GS1l5 strain of <i>Pichia </i>by electroporation. The <i>His⁺ Pichia </i>transformants were anaIysed for the presence of the

insert in the yeast genome by PCR. PCR positive clones of each serotype were grown separately and expression was

induced with 0.5% methanol. The expressed gene product in yeast, <i>Pichia pastoris </i>was characterized by SDS-PAGE

and Western Blot analysis. This is the first report on the production of FMDV structural proteins in yeast.