XK81A1 is a type I epidermal keratin gene expressed in early developmental stages of Xenopus (Jonas et al. 1985). Fusion of the keratin promoter (-5900 to +26) to a human beta globin gene led to fully epidermis-specific accumulation of human globin mRNA and protein when this DNA was injected into fertilized eggs. Further localization of regulatory sequences was accomplished by injecting marked, 5'-deleted keratin gene DNA into fertilized eggs and evaluating tissue specificity of expression. All 5' flanking DNA upstream from -487 could be removed without interfering with keratin gene expression or regulation. These results suggest that the primary mode of regulation of epidermis-specific keratin gene expression is at the level of transcription, and that sequence elements in the 5' flanking region of the keratin gene, between -487 and +26, are responsible for this regulation.