Design and Construction of Multiple-Epitope Recombinant Vaccine Against Foot-and-Mouth Disease Virus Type O

Foot-and-mouth disease (FMD) is an extremely infectious and occasionally fatal viral disease with a rapid onset and a short course that affects cloven-hoofed animals and results in considerable financial losses. For this reason, availability of more reliable and efficient vaccines against Foot-and-mouth disease virus (FMDV) is felt as a necessity. The purpose of this study is to develop a perfectly safe immunogen in order to supplant the usual inactivated vaccine. A tandem repeat multiple-epitope recombinant vaccine against foot-and-mouth disease (FMD) virus (FMDV) type O was developed. In order to develop recombinant epitope vaccines against foot-and-mouth disease virus (FMDV), multiple-epitope recombinant vaccine (rP) comprising various combinations of B cell and T cell epitopes from VP1 capsid protein were synthesized. A 1155-bp gene fragment was amplified by PCR using specific primers. The amplicon was purified and then cloned into expression vector pET-32a. For expression of recombinant protein, pET32a-rP plasmid was transformed into Escherichia coli BL21 (DE3) cells. Recombinant protein was overexpressed with isopropythio-beta-D-galctoside (IPTG). SDSPAGE and DOT blotting were performed for protein determination and verification.The multiple-epitope recombinant was validated by colony-PCR and enzymatic digestion. IPTG with a dosage of 1.0 mmol/L could effectively promote protein expression. SDS-PAGE analysis demonstrated that our constructed prokaryotic expression system pET32a-rP efficiently produces a target recombinant protein with a molecular weight of about 60 kDa. Results showed that prokaryotic expression system is effective in producing the multiple-epitope recombinant immunogen for FDM virus and can be used as a potential method for poly epitope constructs.