Recombinant VP1 Protein of FMD Virus Type O/IRN/2010 as an Immunogenic Peptide Expression System

Background and Aims: Foot and Mouth Disease (FMD) is a highly contagious disease among cloven-hoofed animals. FMD virus has structural and non-structural proteins. Vp1 is the most immunogenic structural peptides of FMD virus, applied for major vaccine studies. The aim of this study was construction of Pet28-VP1 cassette for FMD virus type O/IRN/2010 and expression VP1 peptide as the most immunogenic antigen was the aim of this study. Materials and Methods: FMD virus type O/IRN/2010 was isolated from cattle in Qom, Iran and propagated on IBRS 2 and BHK 21 cell lines. The VP1 gene was amplified using the specific primer pair and RT-PCR technique. The purified PCR product was sub-cloned into the unique BamHI and Xho1 cloning sites to construct the PTZ57R/T -VP1 cassette. The DH 5 strain was transformed with this cassette. The digested VP1 gene was cloned in the digested Pet28 and confirmed using double digestion. Then the Pet28-VP1 construction was transformed in BL21 strain. Results: Expression of VP1 peptide was evaluated by IPTG induction and SDS-PAGE and confirmed using Guinea pig specific polyclonal antibody against FMD virus type O and conjugated rabbit anti Guinea pig antibody-HRP. Also neutralizing antisera titre was protective for vaccinated animals by recombinant VP1 protein.