Mammalian intestinal lactase hydrolyses a variety of beta-glycosides and is processed from a precursor comprising four tandem domains exhibiting sequence similarity, suggestive of multiple duplication events in the evolutionary past. The aim of the present study was to investigate whether genes homologous to the lactase gene exist in animals other than mammals. A reverse transcriptase-PCR strategy using a degenerate mixture of oligonucleotides was developed to search for the presence of transcripts similar in sequence to the mammalian lactase mRNA in the digestive tracts of a bird (the chicken) and an invertebrate (the mussel). Partial cDNAs corresponding to the 3' end of intestinal mRNAs were identified in both animals. In chicken, two cDNAs were isolated, corresponding to 6.5 kb transcripts that used two distinct polyadenylation sites. In mussels, three cDNAs were obtained and classified into two categories. One class of cDNA hybridized to a major mRNA of 3.5 kb and to minor species of 4.5 kb and 6 kb. The second class of cDNA hybridized to a 13 kb transcript, which was approximately twice as large as the mammalian lactase mRNA. Peptide sequences predicted from the chicken and mussel cDNAs confirmed that the proteins are related to mammalian lactase. They also suggested that the chicken protein and one mussel protein are integral molecules anchored in the cell membrane by a C-terminal transmembrane anchor, like lactase. These data provide evidence that proteins phylogenetically related to the mammalian-specific lactase are widespread in the animal kingdom, and that these proteins are expressed in the intestinal tract.