Using the viral DNA extracted from the scab of goat mouth sore as a template, the orf virus (ORFV) 059 gene was amplified by PCR, cloned, sequenced. The 059 gene coding sequence was optimized and synthesized, then linked to vector pET42a(+) and transformed Escherichia coli BL21(DE3). The positive bacteria were cultured in IPTG LB solution, induced expression protein and detected by western blotting. The expression protein was collected with a His column, renatured by the gradient method, and its concentration was measured by Bradford method. BHK21 cells were cultured to a monolayer in a 12 well cell plate. Four treatments for protein F1L, goatpox virus(GTPV), first protein then virus and protein/virus co-incubation were performed. The fluorescence quantitative PCR was used to determine the amount of GTPV adhesion to the cells on incubation 1.0, 6.0 h, to study the effect of F1L protein on the adhesion of GTPV to BHK21 cells. Results showed that 059 gene coding sequence of ORFV Chongqing Shizhu isolate(ORFV-CQsz) was successfully obtained. This gene encodes F1L protein, which has a domain that binds to heparan sulfate receptor on the cell surface, and is the main heparin-binding active protein. The protein carboxy-terminal contains 2 transmembrane regions, which is not easy induced to expression. However, the E. coli transformed prokaryotic expression plasmid constructed using the optimized DNA sequence obtained high-efficient expression of F1L protein induced by IPTG. Western-blot showed that protein had good immunogenicity to ORFV antibody, and the mass concentration of purified refolded F1L protein was about 1.06 mg/mL by Bradford method. The fluorescence quantitative PCR showed the copies of GTPV adhesive BHK21 cells were different under different incubation treatments. F1L protein showed a certain interference effect on GTPV adhesion to BHK21 cells, and it could reduce the copies of virus adhering to cells.