A controversy has existed in the literature for the past several years regarding the number of vertebrate genes encoding the mitochondrial protein that initiates the first step in heme biosynthesis, delta-aminolevulinate synthase [ALAS; succinyl-CoA: glycine C-succinyltransferase (decarboxylating), EC 2.3.1.37]. By analysis of chicken ALAS cDNA clones isolated from both liver and erythroid cells, we show that at least two separate genes encode ALAS mRNAs. These experiments show that (i) two different genes encode the ALAS isozymes found in erythroid and in liver tissues, and (ii) while the product of the erythroid gene (ALASE) is expressed exclusively in erythroid cells, the hepatic form of the enzyme is expressed ubiquitously, suggesting that this is the nonspecific form (ALASN) found in all chicken tissues.