This study aimed to produce prefabricated nerve graft as effective as autologous nerve graft without donor site morbidity for repairing segmental nerve defects. Thirty rats were used and were separated into three groups. In the first group, vein graft excised from jugular vein was sutured to make a bridge between epineural gaps of tibial and peroneal nerve. In the second group, one-quarter of the nerve diameter was incised after excision of the epineurial sheath, and the vein graft was sutured between epineurial gaps. In the third group, the vein graft was sutured between epineurial gaps, and plasmid including vascular endothelial growth factor (VEGF) gene were injected into muscle next to the nerve. Functional and morphological assessments were performed at the end of the 8 weeks. We prefabricated nerve graft by using autologous vein as a conduit material between two intact nerves and by gene therapy, which increases the VEGF level in the medium.