To investigate the molecular phylogenetic status of the Iriomote cat Felis iriomotensis, partial sequences of the mitochondrial 12S rRNA gene (373 bases) and the cytochrome b gene (402 bases) were determined by using the polymerase chain reaction-product direct sequencing technique and then compared with those of seven other feline species. Six Iriomote cats examined in this study showed no intraspecific variation for both genes. The sequence comparisons and the molecular phylogenetic trees indicated that the Iriomote cat is very closely related to the leopard cat Felis bengalensis, which is a widespread species throughout southern and eastern Asia, and that it is reasonable for these two felines to be classified to the same genus. Based on sequence data, the Iriomote cat was estimated to have diverged from the leopard cat arround or less than 0.2 million years ago, and this concurs with the previously reported geological isolation date of the Ryukyu Arc from the Chinese continent. Our results suggest that the geographic barrier has led the fixation of some unique morphological characters into the Iriomote cat population while both the Iriomote cat and the leopard cat still retain very close genetic characters.