Trichophyton rubrum (T. rubrum) is the most common of the superficial fungi. In an effort to better understand the genetic and biochemical makeup of T. rubrum, we generated cDNA libraries from 3 growth stages and used these to isolate 4002 unique expressed sequence tags (ESTs). Sequence comparisons with the Genbank database allowed 1226 of the ESTs to be assigned putative functions or matched with homologs from other organisms. Of the remaining ESTs, 989 were only weakly similar to known sequences and 1787 had no identifiable functions, suggesting that they represent novel genes. We further analyzed the presence of several important genes involved in the growth, metabolism, signal transduction, pathogenesis and drug resistance in T. rubrum. This information was used to newly elucidate important metabolic pathways in T. rubrum. Taken together, our results should form the molecular basis for continued research on the physiological processes and pathogenic mechanisms of T. rubrum, and may lead to a better understanding of fungal drug resistance and identification of new drug targets.