We conducted an electrophoretic survey to examine geographic genetic variation in samples from 17 localities of the Japanese clawed salamander, Onychodactylus japonicus. This species was divided into six genetic groups (N-Tohoku, S-Tohoku, Tsukuba, SW-Honshu, Kinki, and Shikoku) that were largely concordant with clades or subclades recognized in our previous mtDNA study. Although the relationships among these six groups were not clarified, genetic distances between them were not small (mean Nei's D=0.146-0.471). Among these groups, the geographically isolated Tsukuba group was genetically distinct, possibly as a result of population isolation. In a locality of western Honshu, two groups, SW-Honshu and Shikoku, were found to occur sympatrically. Although several presumable hybrid individuals were found, hybridization between these two groups seemed to occur very rarely. These results indicate that the Shikoku group is specifically distinct from the SW-Honshu group, whose range includes the type locality of O. japonicus.