The complete nucleotide sequence and the exon-intron structure of the luciferase gene of the firefly, Luciola lateralis (Coleoptera: Lampyridae) is described. The luciferase gene of the L. lateralis firefly spans 1,971 bp and consists of six introns and seven exons coding for 548 amino acid residues. From samples collected at Boun and Muju, Korea, three isoforms, with identical exon-intron structure, named BU, MJ1 and MJ2, respectively, were obtained. Although the amino acid sequences of MJ1 and MJ2 were identical to those of known luciferase genes of Korean origin, the BU type was novel, differing from each of the MJ1 and MJ2 types by one amino acid. The luciferase sequences of the Korean samples, including those previously revealed, differed only by one-two amino acid residues, but differed by five-six amino acid residues from that of the luciferase gene recorded from specimens from Japan, which suggest genetic divergence has occurred in this species. Phylogenetic analyses using both amino acid and nucleotide sequences further showed that the luciferase gene of the Japanese L. lateralis firefly is genetically distinguishable from that of Korean L. lateralis, suggesting the presence of a genetic subdivision between the L. lateralis dwelling in the Korean Peninsula and Japanese Islands.