1887

Abstract

is a thermally dimorphic fungus, and causes the most prevalent systemic mycosis in Latin America. Infection is initiated by inhalation of conidia or mycelial fragments by the host, followed by further differentiation into the yeast form. Information regarding gene expression by either form has rarely been addressed with respect to multiple time points of growth in culture. Here, we report on the construction of a genomic DNA microarray, covering approximately 25 % of the genome of the organism, and its utilization in identifying genes and gene expression patterns during growth . Cloned, amplified inserts from randomly sheared genomic DNA (gDNA) and known control genes were printed onto glass slides to generate a microarray of over 12 000 elements. To examine gene expression, mRNA was extracted and amplified from mycelial or yeast cultures grown in semi-defined medium for 5, 8 and 14 days. Principal components analysis and hierarchical clustering indicated that yeast gene expression profiles differed greatly from those of mycelia, especially at earlier time points, and that mycelial gene expression changed less than gene expression in yeasts over time. Genes upregulated in yeasts were found to encode proteins shown to be involved in methionine/cysteine metabolism, respiratory and metabolic processes (of sugars, amino acids, proteins and lipids), transporters (small peptides, sugars, ions and toxins), regulatory proteins and transcription factors. Mycelial genes involved in processes such as cell division, protein catabolism, nucleotide biosynthesis and toxin and sugar transport showed differential expression. Sequenced clones were compared with and genome sequences to assess potentially common pathways across species, such as sulfur and lipid metabolism, amino acid transporters, transcription factors and genes possibly related to virulence. We also analysed gene expression with time in culture and found that while transposable elements and components of respiratory pathways tended to increase in expression with time, genes encoding ribosomal structural proteins and protein catabolism tended to sharply decrease in expression over time, particularly in yeast. These findings expand our knowledge of the different morphological forms of during growth in culture.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.027441-0
2009-08-01
2020-01-22
Loading full text...

Full text loading...

/deliver/fulltext/micro/155/8/2795.html?itemId=/content/journal/micro/10.1099/mic.0.027441-0&mimeType=html&fmt=ahah

References

  1. Albuquerque, P., Kyaw, C. M., Saldanha, R. R., Brigido, M. M., Felipe, M. S. & Silva-Pereira, I. ( 2004; ). Pbhyd1 and Pbhyd2: two mycelium-specific hydrophobin genes from the dimorphic fungus Paracoccidioides brasiliensis. Fungal Genet Biol 41, 510–520.[CrossRef]
    [Google Scholar]
  2. Almeida, A. J., Matute, D. R., Carmona, J. A., Martins, M., Torres, I., McEwen, J. G., Restrepo, A., Leão, C., Ludovico, P. & Rodrigues, F. ( 2007; ). Genome size and ploidy of Paracoccidioides brasiliensis reveals a haploid DNA content: flow cytometry and GP43 sequence analysis. Fungal Genet Biol 44, 25–31.[CrossRef]
    [Google Scholar]
  3. Alter, O., Brown, P. O. & Botstein, D. ( 2000; ). Singular value decomposition for genome-wide expression data processing and modeling. Proc Natl Acad Sci U S A 97, 10101–10106.[CrossRef]
    [Google Scholar]
  4. Arraes, F. B., Benoliel, B., Burtet, R. T., Costa, P. L., Galdino, A. S., Lima, L. H., Marinho-Silva, C., Oliveira-Pereira, L., Pfrimer, P. & other authors ( 2005; ). General metabolism of the dimorphic and pathogenic fungus Paracoccidioides brasiliensis. Genet Mol Res 4, 290–308.
    [Google Scholar]
  5. Bagagli, E., Sano, A., Coelho, K. I., Alquati, S., Miyaji, M., de Camargo, Z. P., Gomes, G. M., Franco, M. & Montenegro, M. R. ( 1998; ). Isolation of Paracoccidioides brasiliensis from armadillos (Dasypus noveminctus) captured in an endemic area of paracoccidioidomycosis. Am J Trop Med Hyg 58, 505–512.
    [Google Scholar]
  6. Barelle, C. J., Priest, C. L., Maccallum, D. M., Gow, N. A., Odds, F. C. & Brown, A. J. ( 2006; ). Niche-specific regulation of central metabolic pathways in a fungal pathogen. Cell Microbiol 8, 961–971.[CrossRef]
    [Google Scholar]
  7. Barros, T. F. & Puccia, R. ( 2001; ). Cloning and characterization of a LON gene homologue from the human pathogen Paracoccidioides brasiliensis. Yeast 18, 981–988.[CrossRef]
    [Google Scholar]
  8. Borges-Walmsley, M. I., Chen, D., Shu, X. & Walmsley, A. R. ( 2002; ). The pathobiology of Paracoccidioides brasiliensis. Trends Microbiol 10, 80–87.[CrossRef]
    [Google Scholar]
  9. Brummer, E., Castaneda, E. & Restrepo, A. ( 1993; ). Paracoccidioidomycosis: an update. Clin Microbiol Rev 6, 89–117.
    [Google Scholar]
  10. Cano, M. I., Cisalpino, P. S., Galindo, I., Ramirez, J. L., Mortara, R. A. & da Silveira, J. F. ( 1998; ). Electrophoretic karyotypes and genome sizing of the pathogenic fungus Paracoccidioides brasiliensis. J Clin Microbiol 36, 742–747.
    [Google Scholar]
  11. Cisalpino, P. S., Puccia, R., Yamauchi, L. M., Cano, M. I., da Silveira, J. F. & Travassos, L. R. ( 1996; ). Cloning, characterization, and epitope expression of the major diagnostic antigen of Paracoccidioides brasiliensis. J Biol Chem 271, 4553–4560.[CrossRef]
    [Google Scholar]
  12. Clemons, K. V., Feldman, D. & Stevens, D. A. ( 1989; ). Influence of oestradiol on protein expression and methionine utilization during morphogenesis of Paracoccidioides brasiliensis. J Gen Microbiol 135, 1607–1617.
    [Google Scholar]
  13. Cooke, F. T., Dove, S. K., McEwen, R. K., Painter, G., Holmes, A. B., Hall, M. N., Michell, R. H. & Parker, P. J. ( 1998; ). The stress-activated phosphatidylinositol 3-phosphate 5-kinase Fab1p is essential for vacuole function in S. cerevisiae. Curr Biol 8, 1219–1222.[CrossRef]
    [Google Scholar]
  14. Costa, A. A., Gomez, F. J., Pereira, M., Felipe, M. S., Jesuino, R. S., Deepe, G. S., Jr & de Almeida Soares, C. M. ( 2002; ). Characterization of a gene which encodes a mannosyltransferase homolog of Paracoccidioides brasiliensis. Microbes Infect 4, 1027–1034.[CrossRef]
    [Google Scholar]
  15. da Silva, S. P., Borges-Walmsley, M. I., Pereira, I. S., Soares, C. M., Walmsley, A. R. & Felipe, M. S. ( 1999; ). Differential expression of an hsp70 gene during transition from the mycelial to the infective yeast form of the human pathogenic fungus Paracoccidioides brasiliensis. Mol Microbiol 31, 1039–1050.[CrossRef]
    [Google Scholar]
  16. Derengowski, L. S., Tavares, A. H., Silva, S., Procopio, L. S., Felipe, M. S. & Silva-Pereira, I. ( 2008; ). Upregulation of glyoxylate cycle genes upon Paracoccidioides brasiliensis internalization by murine macrophages and in vitro nutritional stress condition. Med Mycol 46, 125–134.[CrossRef]
    [Google Scholar]
  17. Ebel, F., Schwienbacher, M., Beyer, J., Heesemann, J., Brakhage, A. A. & Brock, M. ( 2006; ). Analysis of the regulation, expression, and localisation of the isocitrate lyase from Aspergillus fumigatus, a potential target for antifungal drug development. Fungal Genet Biol 43, 476–489.[CrossRef]
    [Google Scholar]
  18. Edgar, R., Domrachev, M. & Lash, A. E. ( 2002; ). Gene expression omnibus: NCBI gene expression and hybridization array data repository. Nucleic Acids Res 30, 207–210.[CrossRef]
    [Google Scholar]
  19. Eisen, M. B., Spellman, P. T., Brown, P. O. & Botstein, D. ( 1998; ). Cluster analysis and display of genome-wide expression patterns. Proc Natl Acad Sci U S A 95, 14863–14868.[CrossRef]
    [Google Scholar]
  20. Felipe, M. S., Andrade, R. V., Petrofeza, S. S., Maranhão, A. Q., Torres, F. A., Albuquerque, P., Arraes, F. B., Arruda, M., Azevedo, M. O. & other authors ( 2003; ). Transcriptome characterization of the dimorphic and pathogenic fungus Paracoccidioides brasiliensis by EST analysis. Yeast 20, 263–271.[CrossRef]
    [Google Scholar]
  21. Felipe, M. S., Andrade, R. V., Arraes, F. B., Nicola, A. M., Maranhão, A. Q., Torres, F. A., Silva-Pereira, I., Poças-Fonseca, M. J., Campos, E. G. & other authors ( 2005; ). Transcriptional profiles of the human pathogenic fungus Paracoccidioides brasiliensis in mycelium and yeast cells. J Biol Chem 280, 24706–24714.[CrossRef]
    [Google Scholar]
  22. Ferreira, M. E., Marques Edos, R., Malavazi, I., Torres, I., Restrepo, A., Nunes, L. R., de Oliveira, R. C., Goldman, M. H. & Goldman, G. H. ( 2006; ). Transcriptome analysis and molecular studies on sulfur metabolism in the human pathogenic fungus Paracoccidioides brasiliensis. Mol Genet Genomics 276, 450–463.[CrossRef]
    [Google Scholar]
  23. Fradin, C., Kretschmar, M., Nichterlein, T., Gaillardin, C., d'Enfert, C. & Hube, B. ( 2003; ). Stage-specific gene expression of Candida albicans in human blood. Mol Microbiol 47, 1523–1543.[CrossRef]
    [Google Scholar]
  24. Gadgil, M., Lian, W., Gadgil, C., Kapur, V. & Hu, W. S. ( 2005; ). An analysis of the use of genomic DNA as a universal reference in two channel DNA microarrays. BMC Genomics 6, 66 [CrossRef]
    [Google Scholar]
  25. Goldman, G. H., dos Reis Marques, E., Duarte Ribeiro, D. C., de Souza Bernardes, L. A., Quiapin, A. C., Vitorelli, P. M., Savoldi, M., Semighini, C. P., de Oliveira, R. C. & other authors ( 2003; ). Expressed sequence tag analysis of the human pathogen Paracoccidioides brasiliensis yeast phase: identification of putative homologues of Candida albicans virulence and pathogenicity genes. Eukaryot Cell 2, 34–48.[CrossRef]
    [Google Scholar]
  26. Hayward, R. E., Derisi, J. L., Alfadhli, S., Kaslow, D. C., Brown, P. O. & Rathod, P. K. ( 2000; ). Shotgun DNA microarrays and stage-specific gene expression in Plasmodium falciparum malaria. Mol Microbiol 35, 6–14.[CrossRef]
    [Google Scholar]
  27. Hwang, L., Hocking-Murray, D., Bahrami, A. K., Andersson, M., Rine, J. & Sil, A. ( 2003; ). Identifying phase-specific genes in the fungal pathogen Histoplasma capsulatum using a genomic shotgun microarray. Mol Biol Cell 14, 2314–2326.[CrossRef]
    [Google Scholar]
  28. Izacc, S. M., Gomez, F. J., Jesuino, R. S., Fonseca, C. A., Felipe, M. S., Deepe, G. S. & Soares, C. M. ( 2001; ). Molecular cloning, characterization and expression of the heat shock protein 60 gene from the human pathogenic fungus Paracoccidioides brasiliensis. Med Mycol 39, 445–455.[CrossRef]
    [Google Scholar]
  29. Jesuino, R. S., Azevedo, M. O., Felipe, M. S., Pereira, M. & De Almeida Soares, C. M. ( 2002; ). Characterization of a chaperone ClpB homologue of Paracoccidioides brasiliensis. Yeast 19, 963–972.[CrossRef]
    [Google Scholar]
  30. Jesuino, R. S., Pereira, M., Felipe, M. S., Azevedo, M. O. & Soares, C. M. ( 2004; ). Molecular cloning and characterization of a cDNA encoding the Paracoccidioides brasiliensis L35 ribosomal protein. Med Mycol 42, 217–221.[CrossRef]
    [Google Scholar]
  31. Johannesson, H., Kasuga, T., Schaller, R. A., Good, B., Gardner, M. J., Townsend, J. P., Cole, G. T. & Taylor, J. W. ( 2006; ). Phase-specific gene expression underlying morphological adaptations of the dimorphic human pathogenic fungus, Coccidioides posadasii. Fungal Genet Biol 43, 545–559.[CrossRef]
    [Google Scholar]
  32. Kanetsuna, F. & Carbonell, L. M. ( 1966; ). Enzymes in glycolysis and the citric acid cycle in the yeast and mycelial forms of Paracoccidioides brasiliensis. J Bacteriol 92, 1315–1320.
    [Google Scholar]
  33. Kanetsuna, F., Carbonell, L. M., Moreno, R. E. & Rodriguez, J. ( 1969; ). Cell wall composition of the yeast and mycelial forms of Paracoccidioides brasiliensis. J Bacteriol 97, 1036–1041.
    [Google Scholar]
  34. Kaufman, R. J. ( 1999; ). Stress signaling from the lumen of the endoplasmic reticulum: coordination of gene transcriptional and translational controls. Genes Dev 13, 1211–1233.[CrossRef]
    [Google Scholar]
  35. Levery, S. B., Toledo, M. S., Suzuki, E., Salyan, M. E., Hakomori, S., Straus, A. H. & Takahashi, H. K. ( 1996; ). Structural characterization of a new galactofuranose-containing glycolipid antigen of Paracoccidioides brasiliensis. Biochem Biophys Res Commun 222, 639–645.[CrossRef]
    [Google Scholar]
  36. Levery, S. B., Toledo, M. S., Straus, A. H. & Takahashi, H. K. ( 1998; ). Structure elucidation of sphingolipids from the mycopathogen Paracoccidioides brasiliensis: an immunodominant β-galactofuranose residue is carried by a novel glycosylinositol phosphorylceramide antigen. Biochemistry 37, 8764–8775.[CrossRef]
    [Google Scholar]
  37. Levery, S. B., Toledo, M. S., Doong, R. L., Straus, A. H. & Takahashi, H. K. ( 2000; ). Comparative analysis of ceramide structural modification found in fungal cerebrosides by electrospray tandem mass spectrometry with low energy collision-induced dissociation of Li+ adduct ions. Rapid Commun Mass Spectrom 14, 551–563.[CrossRef]
    [Google Scholar]
  38. Lorenz, M. C. & Fink, G. R. ( 2001; ). The glyoxylate cycle is required for fungal virulence. Nature 412, 83–86.[CrossRef]
    [Google Scholar]
  39. Lorenz, M. C. & Fink, G. R. ( 2002; ). Life and death in a macrophage: role of the glyoxylate cycle in virulence. Eukaryot Cell 1, 657–662.[CrossRef]
    [Google Scholar]
  40. Lorenz, M. C., Bender, J. A. & Fink, G. R. ( 2004; ). Transcriptional response of Candida albicans upon internalization by macrophages. Eukaryot Cell 3, 1076–1087.[CrossRef]
    [Google Scholar]
  41. Mackinnon, J. E. ( 1970; ). On the importance of South American blastomycosis. Mycopathol Mycol Appl 41, 187–193.[CrossRef]
    [Google Scholar]
  42. Manly, K. F., Nettleton, D. & Hwang, J. T. ( 2004; ). Genomics, prior probability, and statistical tests of multiple hypotheses. Genome Res 14, 997–1001.[CrossRef]
    [Google Scholar]
  43. Manocha, M. S. ( 1980; ). Lipid composition of Paracoccidioides brasiliensis: comparison between the yeast and mycelial forms. Sabouraudia 18, 281–286.[CrossRef]
    [Google Scholar]
  44. Marques, E. R., Ferreira, M. E., Drummond, R. D., Felix, J. M., Menossi, M., Savoldi, M., Travassos, L. R., Puccia, R., Batista, W. L. & other authors ( 2004; ). Identification of genes preferentially expressed in the pathogenic yeast phase of Paracoccidioides brasiliensis, using suppression subtraction hybridization and differential macroarray analysis. Mol Genet Genomics 271, 667–677.
    [Google Scholar]
  45. Mattar-Filho, R., Azevedo, M. O., Pereira, M., Jesuino, R. S., Salem-Izacc, S. M., Brito, W. A., Gesztesi, J. L., Soares, R. B., Felipe, M. S. & Soares, C. M. ( 1997; ). Expression of glycoprotein gp43 in stage-specific forms and during dimorphic differentiation of Paracoccidioides brasiliensis. J Med Vet Mycol 35, 341–345.[CrossRef]
    [Google Scholar]
  46. McCullough, M. J., DiSalvo, A. F., Clemons, K. V., Park, P. & Stevens, D. A. ( 2000; ). Molecular epidemiology of Blastomyces dermatitidis. Clin Infect Dis 30, 328–335.[CrossRef]
    [Google Scholar]
  47. Mo, C., Valachovic, M. & Bard, M. ( 2004; ). The ERG28-encoded protein, Erg28p, interacts with both the sterol C-4 demethylation enzyme complex as well as the late biosynthetic protein, the C-24 sterol methyltransferase (Erg6p). Biochim Biophys Acta 1686, 30–36.[CrossRef]
    [Google Scholar]
  48. Montoya, A. E., Alvarez, A. L., Moreno, M. N., Restrepo, A. & McEwen, J. G. ( 1999; ). Electrophoretic karyotype of environmental isolates of Paracoccidioides brasiliensis. Med Mycol 37, 219–222.[CrossRef]
    [Google Scholar]
  49. Nunes, L. R., Costa de Oliveira, R., Leite, D. B., da Silva, V. S., dos Reis Marques, E., da Silva Ferreira, M. E., Ribeiro, D. C., de Souza Bernardes, L. A., Goldman, M. H. & other authors ( 2005; ). Transcriptome analysis of Paracoccidioides brasiliensis cells undergoing mycelium-to-yeast transition. Eukaryot Cell 4, 2115–2128.[CrossRef]
    [Google Scholar]
  50. Oefner, P. J., Hunicke-Smith, S. P., Chiang, L., Dietrich, F., Mulligan, J. & Davis, R. W. ( 1996; ). Efficient random subcloning of DNA sheared in a recirculating point-sink flow system. Nucleic Acids Res 24, 3879–3886.[CrossRef]
    [Google Scholar]
  51. Paris, S., Duran-Gonzalez, S. & Mariat, F. ( 1985; ). Nutritional studies on Paracoccidioides brasiliensis: the role of organic sulfur in dimorphism. Sabouraudia 23, 85–92.[CrossRef]
    [Google Scholar]
  52. Patel, J. B., Batanghari, J. W. & Goldman, W. E. ( 1998; ). Probing the yeast phase-specific expression of the CBP1 gene in Histoplasma capsulatum. J Bacteriol 180, 1786–1792.
    [Google Scholar]
  53. Rappleye, C. A. & Goldman, W. E. ( 2006; ). Defining virulence genes in the dimorphic fungi. Annu Rev Microbiol 60, 281–303.[CrossRef]
    [Google Scholar]
  54. Restrepo, A. & Jimenez, B. E. ( 1980; ). Growth of Paracoccidioides brasiliensis yeast phase in a chemically defined culture medium. J Clin Microbiol 12, 279–281.
    [Google Scholar]
  55. Richie, D. L., Hartl, L., Aimanianda, V., Winters, M. S., Fuller, K. K., Miley, M. D., White, S., McCarthy, J. W., Latgé, J. P. & other authors ( 2009; ). A role for the unfolded protein response (UPR) in virulence and antifungal susceptibility in Aspergillus fumigatus. PLoS Pathog 5, e1000258 [CrossRef]
    [Google Scholar]
  56. Rooney, P. J. & Klein, B. S. ( 2002; ). Linking fungal morphogenesis with virulence. Cell Microbiol 4, 127–137.[CrossRef]
    [Google Scholar]
  57. Rozen, S. & Skaletsky, H. ( 2000; ). Primer3 on the WWW for general users and for biologist programmers. Methods Mol Biol 132, 365–386.
    [Google Scholar]
  58. San Blas, G. & San Blas, F. ( 1977; ). Paracoccidioides brasiliensis: cell wall structure and virulence. Mycopathologia 62, 77–86.[CrossRef]
    [Google Scholar]
  59. San-Blas, G. & Vernet, D. ( 1977; ). Induction of the synthesis of cell wall α-1,3-glucan in the yeastlike form of Paracoccidioides brasiliensis strain IVIC Pb9 by fetal calf serum. Infect Immun 15, 897–902.
    [Google Scholar]
  60. Schobel, F., Ibrahim-Granet, O., Avé, P., Latgé, J. P., Brakhage, A. A. & Brock, M. ( 2007; ). Aspergillus fumigatus does not require fatty acid metabolism via isocitrate lyase for development of invasive aspergillosis. Infect Immun 75, 1237–1244.[CrossRef]
    [Google Scholar]
  61. Sebghati, T. S., Engle, J. T. & Goldman, W. E. ( 2000; ). Intracellular parasitism by Histoplasma capsulatum: fungal virulence and calcium dependence. Science 290, 1368–1372.[CrossRef]
    [Google Scholar]
  62. Servant, G., Pennetier, C. & Lesage, P. ( 2008; ). Remodeling yeast gene transcription by activating the Ty1 long terminal repeat retrotransposon under severe adenine deficiency. Mol Cell Biol 28, 5543–5554.[CrossRef]
    [Google Scholar]
  63. Sprague, G. F. & Cronan, J. E. ( 1977; ). Isolation and characterization of Saccharomyces cerevisiae mutants defective in glycerol catabolism. J Bacteriol 129, 1335–1342.
    [Google Scholar]
  64. Stover, E. P., Schar, G., Clemons, K. V., Stevens, D. A. & Feldman, D. ( 1986; ). Estradiol-binding proteins from mycelial and yeast-form cultures of Paracoccidioides brasiliensis. Infect Immun 51, 199–203.
    [Google Scholar]
  65. Talaat, A. M., Howard, S. T., Hale, W., 4th, Lyons, R., Garner, H. & Johnston, S. A. ( 2002; ). Genomic DNA standards for gene expression profiling in Mycobacterium tuberculosis. Nucleic Acids Res 30, e104 [CrossRef]
    [Google Scholar]
  66. Tavares, A. H., Silva, S. S., Dantas, A., Campos, E. G., Andrade, R. V., Maranhão, A. Q., Brígido, M. M., Passos-Silva, D. G., Fachin, A. L. & other authors ( 2007; ). Early transcriptional response of Paracoccidioides brasiliensis upon internalization by murine macrophages. Microbes Infect 9, 583–590.[CrossRef]
    [Google Scholar]
  67. Thirach, S., Cooper, C. R., Jr & Vanittanakom, N. ( 2008; ). Molecular analysis of the Penicillium marneffei glyceraldehyde-3-phosphate dehydrogenase-encoding gene (gpdA) and differential expression of gpdA and the isocitrate lyase-encoding gene (acuD) upon internalization by murine macrophages. J Med Microbiol 57, 1322–1328.[CrossRef]
    [Google Scholar]
  68. Thorstenson, Y. R., Hunicke-Smith, S. P., Oefner, P. J. & Davis, R. W. ( 1998; ). An automated hydrodynamic process for controlled, unbiased DNA shearing. Genome Res 8, 848–855.
    [Google Scholar]
  69. Toledo, M. S., Levery, S. B., Straus, A. H., Suzuki, E., Momany, M., Glushka, J., Moulton, J. M. & Takahashi, H. K. ( 1999; ). Characterization of sphingolipids from mycopathogens: factors correlating with expression of 2-hydroxy fatty acyl (E)-Δ3-unsaturation in cerebrosides of Paracoccidioides brasiliensis and Aspergillus fumigatus. Biochemistry 38, 7294–7306.[CrossRef]
    [Google Scholar]
  70. Wang, Z. Y., Thornton, C. R., Kershaw, M. J., Debao, L. & Talbot, N. J. ( 2003; ). The glyoxylate cycle is required for temporal regulation of virulence by the plant pathogenic fungus Magnaporthe grisea. Mol Microbiol 47, 1601–1612.[CrossRef]
    [Google Scholar]
  71. Wanke, B. & Londero, A. T. ( 1994; ). Epidemiology and paracoccidioidomycosis infection. In Paracoccidioidomycosis, pp. 109–120. Edited by M. Franco, C. da Silva Lacaz, A. Restrepo-Moreno & G. Del Negro. Boca Raton, FL: CRC Press.
  72. Williams, B. A., Gwirtz, R. M. & Wold, B. J. ( 2006; ). Genomic DNA as a general cohybridization standard for ratiometric microarrays. Methods Enzymol 410, 237–279.
    [Google Scholar]
  73. Yamamoto, A., DeWald, D. B., Boronenkov, I. V., Anderson, R. A., Emr, S. D. & Koshland, D. ( 1995; ). Novel PI(4)P 5-kinase homologue, Fab1p, essential for normal vacuole function and morphology in yeast. Mol Biol Cell 6, 525–539.[CrossRef]
    [Google Scholar]
  74. Zaugg, C., Monod, M., Weber, J., Harshman, K., Pradervand, S., Thomas, J., Bueno, M., Giddey, K. & Staib, P. ( 2009; ). Gene expression profiling in the human pathogenic dermatophyte Trichophyton rubrum during growth on proteins. Eukaryot Cell 8, 241–250.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.027441-0
Loading
/content/journal/micro/10.1099/mic.0.027441-0
Loading

Data & Media loading...

Supplements

[PDF file](43 KB)

PDF

[PDF file](653 KB)

PDF

List of primers for PCR controls for known P. brasiliensis genes spotted on the microarray as well as primers and probes for the genes selected for real-time RT-PCR [PDF file](156 KB)

PDF

[PDF file](84 KB)

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error