Verticillium dahliae is a principal pathogen causing verticillium wilt in Solanaceae crops. StoVe1 is a gene resisting to verticillium wilt isolated from Solanum torvum. In order to generate resistant tobacco plants, StoVe1 was inserted in the orientation behind CaMV 35S promoter of vector pGS and this construct was introduced into tobacco by Agrobacterium-mediated transformation. A total of 12 kanamycin-resistant plants were generated and 7 independent transgenic lines were identified by PCR analysis. Quantitative RT-PCR analysis showed that the levels of StoVe1 transcript in transgenic lines were up to 2-6 fold higher than in the control. Anti-fungal assay indicated that the protein extract of transgenic tobacco lines showed strong inhibition activity to V. dahliae, 2 fold or higher compared to control plants. This result reveals that StoVe1, as a V. dahliae resistance gene, has an application potential in plant breeding for verticillium wilt resistance.