A highly repeated interspersed sequence (OLR1) was isolated from a genomic DNA library of the Medaka, Oryzias latipes. The OLR1 was about 160 base pairs (bp) in length. As judged from the results of colony hybridization experiments, OLR1 is one of the major repeated DNA sequences in the Medaka genome and is present in every 136 kb on average. Results of Southern and colony-hybridization analyses indicate that OLR1 is a small interspersed repetitive element (SINE). OLR1-related sequences were conserved in other three species (O. luzonensis, O. curvinotus, and O. mekongnensis) within the genus Oryzias as a repetitive sequence. These results lend support at the DNA level to the hypothesis that these four species form one group in the genus Oryzias, as has been suggested from an analysis of their karyotypes (Magtoon and Uwa, '85, Proc. Jpn. Acad., Ser. B, 61:157-160).