The enzyme 17beta-hydroxysteroid dehydrogenase (17betaHSD) interconverts 17-ketosteroids and 17beta-hydroxysteroids. Five isoforms of the enzyme have been identified in the human, two of which (types 1 and 3) have been shown to catalyse the reduction reaction preferentially. The cDNA encoding mouse 17betaHSD type 3 was isolated from testis cDNA using the RACE technique and primers based on the human sequence. The mouse protein is 305 amino acids in length which is five short of the human protein with four of these amino acids missing at the N-terminus. The predicted amino acid sequence is 72.5% identical and 94.8% similar to the human sequence. Tissue distribution of mRNA encoding both types 1 and 3 17betaHSD was studied using reverse transcription and the polymerase chain reaction (RT-PCR). Highest levels of type 1 mRNA were found in the ovary whereas highest levels of type 3 were in the testis. All other tissues tested contained mRNA encoding both isoforms of the enzyme although levels were markedly lower than in the gonads.