Abstract: The anti-foot-and-mouth disease virus(FMDV) single-chain Fv(scFv) gene was constructed by DNA recombinant technology.The 1C7V_H and V_L genes were assembled into scFv gene with a linker sequence(Gly_4Ser)_3 by splicing overlop extension and scFv gene was amplified by PCR.The scFv gene was subcloned into phagemid vector pCANTAB 5E.The phage scFv displaying library was constructed by male Escherichia coli TG1 transfected with resultant phagemid pCANTAB 5E-scFv.The scFv positive clone was obtained after the phage scFv displaying library was rescued by M13KO7 and biopanned with FMDV antigen for 3 rounds.Escherichia coli BH2151 was transfected with the positive clone and scFv protein was expressed by Escherichia coli BH2151 through the inducement of IPTG.The results of ELISA show that phage-scFv and soluble scFv,which were expressed by the selected scFv gene,have high affinity and specificity to FMDV.