For example, transmembrane

proteins involved in the transport of metallic ions appear to play an important role in microbial pathogenesis [51] as demonstrated in the Cu2+-ATPase mutants of EPZ015938 ic50 Listeria Vorinostat research buy monocytogenes [52] and Criptococcus neoformans [53] that show reduced virulence. In the latter case, the Δvph1 mutant did not display laccase activity, which is an essential virulence factor of this pathogen [53]. Moreover, an ATP-binding cassette (ABC) transporter listed in Table 2 is overexpressed in mycelia cultured in keratin, suggesting its involvement in T. rubrum pathogenicity. In addition, the strain carrying a disrupted version of this MDR gene (ΔTruMDR2) showed low infectious capability characterized by reduced growth of T. rubrum on human nails [40]. Conclusions We identified 575 novel ESTs and obtained new molecular data related to T. rubrum growth, pH and carbon source signaling, and stress responses to antifungal challenges. It is clear that additional studies are necessary to define the functioning of whole genes and fully understand the regulation of these complex adaptive responses. However, the various ESTs identified in this work provide new insights into different aspects of T. rubrum biology,

revealing new sources for functional genome analysis. T. rubrum genes that encode putative proteins similar CRT0066101 cost to virulence factors described for other fungi were among the ESTs identified. The transcriptional profile also suggested that several genes could function in environmental

stress responses. Thus, our study can help to better understand the molecular mechanisms of the adaptive responses possibly involved in dermatophyte infection and antifungal resistance. Methods Strains and culture conditions The H6 (ATCC MYA-3108) and F6 mutant (a fluconazole-resistant strain isolated in our laboratory) strains of T. rubrum were cultured on Sabouraud dextrose agar plates (SDA) as described earlier [54]. Phosphatidylethanolamine N-methyltransferase The F6 cultures were supplemented with fluconazole (200 μg/mL). Conidia from these strains were used to construct the cDNA (library 1) or were inoculated in Sabouraud dextrose broth (SDB) and incubated for 72 h at 28°C on an orbital shaker at 180 rpm. The resulting mycelia were aseptically transferred to the desired culture media, and these were used to construct each of the SSH libraries. Construction of the libraries One cDNA library (Library 1) and nine SSH libraries (Libraries 2 to 10) were constructed. The SSH libraries were performed between the tester and driver DNA, with the cDNA population containing the differentially expressed transcripts being the tester, and the reference cDNA (control) being the driver.