The nucleotide (nt) sequence of the dihydrofolate reductase (DHFR)-encoding gene (DFR1) of Candida albicans was determined. The gene contains an open reading frame of 576 nt, coding for a protein of 192 amino acid (aa) residues (calculated M(r) 22,222), that is 38.5 and 31% similar to the Saccharomyces cerevisiae and human enzymes, respectively. The first 36 residues, at the N terminus, of the deduced aa sequence are identical to those determined by sequencing of the purified enzyme from C. albicans. Putative transcription start points were also determined. Restriction-fragment-length polymorphism analysis of the DFR1 chromosomal region suggests the presence of a single copy of the gene per haploid genome and shows a limited variability among the different C. albicans strains tested.