A cDNA clone encoding 10-formyltetrahydrofolate dehydrogenase (10-FTHFDH) was isolated from a human fetal liver cDNA library. It contained the open reading frame of 2,709 base pairs and predicted a protein comprising 902 amino acids with a calculated molecular weight of 98,700 Da. The deduced protein showed about 93.6% homology (90.5% identity, 3.1% favored substitutions) when compared with rat 10-FTHFDH. The distribution of 10-FTHFDH transcript in various human tissues was studied by Northern blot analysis using poly(A+) RNAs from different tissues. The 10-FTHFDH transcript with an approximate size of 2.7 kb was mainly expressed in human kidney, skeletal muscle, and liver and rarely expressed in other tissues.