Eclosion hormone (EH), one of the neuropeptides, serves as a critical factor in the control of ecdysis behavior and development in insects. The Asian corn borer, Ostrinia furnacalis (Osf) is one of the major agricultural pests in Asia. In this study, the molecular characterizations and tissue expression of the eclosion hormone gene in O. furnacalis were elucidated. Using degenerate primers and RT-PCR method, the middle part of the cDNA of EH gene from O. furnacalis was cloned and sequenced. The full-length cDNA of EH gene was then obtained with rapid amplification of cDNA ends (RACE) method. The full Osf-EH cDNA is 986 bp (GenBank accession no. DQ668369), containing a 267 bp open reading frame encoding an 88-amino acid preprohormone. After translation,  the preprohormone was cleavaged to a　hydrophobic 26-amino acids signal peptide and a 62-amino acids mature peptide. The Osf-EH shows high identity with those from Manduca sexta (79.5%), Helicoverpa armigera (77.3%) and Bombyx mori (67.0%), but low identity with that of Drosophila melanogaster (45.5%). RT-PCR analysis showed that the EH mRNA only expressed in the brain, and was not be detected in other neural tissues, such as suboesophageal ganglion, thoracic ganglion, abdominal ganglion, and non-neural tissues, such as midgut, fat body and epidermis.