Recombinant VP1 protein of FMD virus type O/IRN/2010 as an immunogenic peptide expression system on guinea pig

Foot and Mouth Disease (FMD) is 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. 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. The FMD virus type O/IRN/2010 was isolated from cattle in Qom, Iran and propagated on IBRS2 and BHK21 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 DH5&alpha 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. Expression of VP1 peptide was evaluated by IPTG induction and SDS-PAGE and confirmed by 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. Since the isolation of new FMD virus strains in different geographical locations and expression of VP1 peptide can be used in emergency and control settings as a recombinant vaccine in the same area. Therefore the Pet28-VP1 cassette was constructed in this study is a good candidate for preparation of peptide vaccine against FMD virus type O/IRN/2010 in the next future.