Gyrate atrophy of the choroid and retina is an autosomal recessive disease associated with reduced or absent ornithine aminotransferase (OAT) activity. To approach the defect in OAT at the molecular level, we have cloned a cDNA for the mRNA encoding the OAT precursor from human liver. The clone contains the complete coding region of 1317 nucleotides along with 44 nucleotides of 5' and 654 nucleotides of 3' untranslated sequences. When used to probe genomic DNA, the OAT cDNA did not detect any evidence of gene deletion or rearrangement in patients with gyrate atrophy. The cDNA hybridizes to a 2.15-kb RNA species in liver, fibroblasts, and lymphoblasts. The size and approximate amount of this mRNA is not altered in fibroblasts and/or lymphoblasts of seven gyrate atrophy patients who display a 25- to 100-fold reduction in OAT activity. Our results suggest the defect in these individuals may be caused by a subtle sequence alteration in the mRNA that does not affect its apparent size.