Delta-like 1 homolog (Dlk1) is a paternally expressed imprinted gene in mammals, regulating development and differentiation of adipose and muscle. The Dlk1 genes of the quail and turkey were cloned and analyzed in their properties of amino acid sequences, alternative splicing, and genetic distances from other species. In addition, because Dlk1 is located in the cluster of up to 10 imprinted genes in mammals, the genomic structure of the cluster was investigated in the chicken. Furthermore, the imprinting status of the avian Dlk1 gene was also determined here. The numbers of coding sequences of the quail and turkey Dlk1 were the same as chicken Dlk1 in nucleotide (1,161 bp) and amino acid (386 amino acids) sequences. The amino acid similarities were more than 96% with predicted conserved domains including the signal sequence, 6 epidermal growth factor-like domains, and a transmembrane domain. As in the chicken, the alternative splicing of Dlk1 transcripts was not observed in the turkey and quail. Phylogenetic analysis revealed that the chicken and turkey Dlk1 were closer than the chicken and quail. Comparative analysis of the gene clusters containing the Dlk1 gene revealed that Yy1, Wars, Wdr25, Begain, Dlk1, Dio3, and Ppp2r5c were found in the cluster of the chicken genome, but 3 genes (Meg3, Rtl1, and Meg8) between Dlk1 and deiodinase, iodothyronine, type III (Dio3) were not found. Several SNP in the genomic DNA sequences of the fifth exon were identified in chickens and quail. Sequencing analysis of reverse transcription-PCR products of Dlk1 revealed that adipose and muscle from chickens and quail heterozygous for these SNP produce Dlk1 transcripts from both alleles, demonstrating biallelic expression of Dlk1 in the avian species. These results clearly demonstrate that avian Dlk1 is not imprinted and its expression might be regulated in a different manner from mammals.