Abstract: Foot-and-Mouth Disease Virus (FMDV) has the high mutation rates and exhibits an important potential for adaptations. The new variants will be generated upon environment change to adapt new environmental conditions, these variants contained one or several amino acid replacements within the capsid protein, including the RGD cell receptor-binding motif. Following serial passages of FMDV Asia1/JS/China/05 isolates under different host condition, the viruses which contained an RSD and RDD sequence in the cell receptorbinding site were generated. To study the effect of single amino acid replacements within RDD receptor-binding motif on production of infectious virus particle, we constructed type Asia 1 FMDV full-length cDNA clone pFMDV-RSD by using over-lap PCR, based on FMDV Asia1/JS/China/05 full-length infectious cDNA clone- BHK-21 cells were cotransfected with linearized recombinant plasmids and plasmids expressing T7 RNA polymerase Apparent CPE were observed after 56 h incubation. Furthermore, the rescued FMDV were verified by RT-PCR, IFA, electron microscope and sulk mice pathogenicity analysis. These results suggested that the infectious FMDV were acquired, and one amino acid replacement in the cell receptorbinding did not affect the virus viability. This study lays a foundation for further study of biology characteristic of rescued virus with RSD receptor recognition site.