cerevisiae and Aspergillus fumigatus) revealed the presence of two distinct regions. The one

located at the 5′- region showed high homology with the Spe genes, whereas the one present at the 3′-region was homologous to the Sdh genes; both were linked through a region of approximately 60 nucleotides without RO4929097 mouse homology (not shown). As expected, the alignment of amino acid sequences encoded by these genes showed the same pattern of homology, demonstrating the high preservation of the gene in the Basidiomycota (not shown). With these data we designed degenerate primers to be used for PCR amplification of the chimeric genes. The forward primer was selected at the 3′-end of the region with homology to Spe, and the reverse primer was designed from the homologous region

at the 5′-end of the Sdh, in such a way that the amplification fragment covered the nonhomologous region that separates both coding regions (see Fig. 1a). Using the PCR conditions described above and AZD2014 the designed degenerate primers, it was possible to amplify DNA fragments of the predicted size from genomic DNA of all the Basidiomycota species tested (see Materials and methods), whose genomes have been sequenced or not, that represented the three subphyla from Basidiomycota. The size of the fragments (around 1300 bp) coincided with the expected values. On the other hand, and as expected, no such amplification occurred when DNA from Ascomycota or Zygomycota species was used as template (Fig. 1b). The PCR products corresponding to the Basidiomycota species analyzed in this work were sequenced. Alignment of the encoded sequences revealed their high conservation (Fig. 2). Additionally, the encoded sequences

of the amplified fragments from Basidiomycota species whose genomes had been previously sequenced were compared with those existing in their corresponding data banks. The results obtained confirmed the fidelity of the PCR amplification Mannose-binding protein-associated serine protease (Table 1). The differences observed can be explained by the fact that different isolates were used in these studies. The sequences of the fragments were deposited in GenBank, with the following accession numbers: Ustilago cynodontis, FN646089; Tilletia foetida, FN646090; Bjerkandera adusta, FN646091; Rhizoctonia solani, FN822770; Schizophyllum commune, FN822771; Ustilago hordei, FN822772; Ustilago maydis, FN822773; Coprinus cinerea, FN822774; Pleurotus ostreatus, FN822775; Ganoderma lucidum, FN822776; Agaricus bisporus, FN827330; and Ganoderma sp., FN827329. The sequences of the regions corresponding to the fragments amplified by PCR from the Spe-Sdh genes obtained in this study, and those reported in the databases, were used for the construction of a phylogenetic tree. The results obtained showed the phylogenetic relationship (Fig.