The antibody induced by non-structure of foot-and-mouth disease virus (FMDV) could be used as serological marker to distinguish between nature infection and vaccinated animal. This year, we focused on the non-structure gene of FMDV to design primer and amplified the target gene by the RT-PCR method. The RT-PCR products were inserted into expression vector and were cloned into the prokaryotic and eukaryotic cell to acquire the recombinant protein. The complete target gene were constructed and transformed to the E.coli and the Baculovirus system. A great deal of recombinant protein was produced by the expressing system. The recombinant protein was confirmed on the Western blot assay and it showed the same as function like the natural non-structure protein produced by FMDV. High titer of polyclonal and monoclonal antibodies against the non-structural protein have been prepared already. We have tried to build up a manufacturing process for pen-side diagnostic set for FMD by using recombinant non-structure protein.