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Hepatic and intestinal expressed sequence tags (ESTs) of lactating German Holstein and Charolais cows were used to identify and map genes potentially involved in the regulation of metabolic processes in cattle. Seventeen ESTs were homologous to human gene sequences and six ESTs identified unknown coding sequences in human genomic DNA clones. A cattle–hamster somatic hybrid cell panel and cattle–hamster 5000 rad whole genome radiation hybrid (WGRH) panel were used to assign the ESTs in the bovine genome. Fifteen of the mapped genes and the six ESTs represent new assignments in the established cattle WGRH5000 map. Calculation of the radiation hybrid (RH) mapping data was performed with the software RHMAPPER. The new loci contribute to the completion of the bovine transcript maps and their comparative assignments increase our knowledge about genome evolution between cattle and human.