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The primary structure of the α-subunit of the adenylate cyclase-inhibiting G-protein (g i ) has been deduced from the nucleotide sequence of cloned DNA complementary to the bovine cerebral mRNA encoding the polypeptide. A much higher degree of amino acid sequence homology is observed between the α-subunits of G i and transducin (68%) than between those of G i and the adenylate cyclase-stimulating G-protein (G s ) (43%) or between those of transucin and G s (42%).