We have isolated and sequenced cDNA clones that code for rat and human NADPH-dependent cytochrome P-450 reductase. The cDNA coding for the human protein was used to analyse, by Southern blot hybridization, DNA isolated from a panel of 8 independent human-rodent somatic cell hybrids. The results indicate that cytochrome P-450 reductase is encoded by a single gene (POR) located on human chromosome 7(pter-q22). Analysis of human metaphase chromosomes by hybridization in situ confirmed the results and refined the localization to 7q11.2. Northern blot hybridization revealed that in human liver the expression of the gene varies by less than 3-fold between different individuals.