Naturally occurring regulatory variation is a source of genetic variability that is well documented but poorly understood. Two members of the Hawaiian picture-winged Drosophila, D. affinidisjuncta and D. hawaiiensis, display markedly different levels of alcohol dehydrogenase (alcohol: NAD+ oxidoreductase, EC 126.96.36.199) in the larval midgut and Malpighian tubules. To analyze the regulation of the alcohol dehydrogenase genes from these two species, their homologous alcohol dehydrogenase genes were cloned and introduced, via P element-mediated transformation, into the germ line of Drosophila melanogaster. Histochemical and electrophoretic analyses of larval transformants demonstrate that major differences in the tissue-specific levels of alcohol dehydrogenase production are characteristic of the alcohol dehydrogenase genes themselves. While these results do not directly address possible species-specific differences in the tissue distribution of trans-acting regulatory components, they indicate that demonstrable differences in cis-dominant regulatory information are sufficient to account for the observed regulatory variation.