Abstract: A pair of primer was designed according to the 2C gene sequence of footandmouth disease virus (FMDV) for amplification of a fragment including 174 bp 5′-end and 279 bp 3′-end of 2C gene， which encoded abundant B-cell epitopes of 2C protein. The amplified fragment was inserted into pET-30a plasmid (Novagen) via two unique restriction sites of NcoⅠand SalⅠ. Open reading frame of target gene was confirmed correctly inserted into the positive recombinant plasmid by sequencing， the recombinant plasmid was transformed into the host strain bacteria BL21(DE3)pLys for protein expression. After inducing by IPTG at 37 ℃ for five hours， the expressed product was analyzed by SDS-PAGE and Western Blotting. The results revealed that the target gene fragment of 2C had been expressed successfully. The product is a 23 kD fusion protein and can react with sera derived from FMDV infected animal. It would provide an useful antigen for establishment of enzyme linked immunetransfer blot (EITB) diagnostic method， which can be used for differentiation of the FMDV infected animals from the vaccinated animals.