A recombinant Rabies virus (RV) expressing firefly luciferase (rRV-luc) was generated by an improved reverse genetics system. Its biological properties were compared with those of the parental RV. The rRV-luc grew in BHK-21 cells similarly to RV, but its virulence for mice was weaker as shown by the lower infectious titers in brain. Rising infectious titers of rRV-luc during its passaging in BHK-21 cells indicated a virus adaptation, while the luciferase (luc) expression was stable. These results suggest that the recombinant RV carrying luc gene might prove a useful tool for further analysis of pathogenesis of RV in small animal models.