cDNA clones coding for glucagon were isolated from a chicken pancreas cDNA library, and the nucleotide and amino acid sequences were determined. The amino acid sequence of chicken glucagon was HSQGTFTSDYSKYLDSRRAQDFVQWLMST, which was contained in the 151-amino acid long precursor, being preceded by a signal sequence and an amino-terminal peptide (NH2-peptide) and followed by an intervening peptide and a glucagon-like peptide I (GLP-I). Chicken preproglucagon, however, lacked GLP-II and intervening peptide II which have been shown to be contained in mammalian glucagon precursors.