Vibrio is one of the most common bacterial groups in marine environments.Molecular basis of pathogenesis of Vibrio keeps little known mainly due to limited genetic tools for functional research of genes in most pathogenic Vibrio species.In this study,we developed universal genetic tools for rapid and efficient deletion mutation in Vibrio species based on suicide T-Vectors carrying a novel counterselectable marker,vmi480.We explored two uncharacterized genes,vmi480 and vmi470,in a genomic island from Vibrio mimicus VM573,and confirmed that vmi480 and vmi470 constituted a two-component toxin-antitoxin system through deletion and expression of vmi480 and vmi470.