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Modular Gene Over-expression Strategies for Candida albicans

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Host-Fungus Interactions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 845))

Abstract

Over-expression is a valid functional genomics approach to characterise genes of unknown function on a genome-wide scale. Strains are engineered to over-express a specific gene and the resulting gain-of-function phenotype assessed. Here, we describe the strategy we are adopting to synthesise a Candida albicans ORFeome collection and the options available to create over-expressing strains from this collection.

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References

  1. Sopko, R., Huang, D., Preston, N., Chua, G., Papp, B., Kafadar, K., Snyder, M., Oliver, S. G., Cyert, M., Hughes, T. R., Boone, C., and Andrews, B. (2006) Mapping pathways and phenotypes by systematic gene overexpression, Mol Cell21, 319–330.

    Article  CAS  Google Scholar 

  2. Chua, G., Morris, Q. D., Sopko, R., Robinson, M. D., Ryan, O., Chan, E. T., Frey, B. J., Andrews, B. J., Boone, C., and Hughes, T. R. (2006) Identifying transcription factor functions and targets by phenotypic activation, Proc Natl Acad Sci USA103, 12045–12050.

    Article  CAS  Google Scholar 

  3. Hiller, D., Sanglard, D., and Morschhauser, J. (2006) Overexpression of the MDR1 gene is sufficient to confer increased resistance to toxic compounds in Candida albicans, Antimicrob Agents Chemother50, 1365–1371.

    Article  CAS  Google Scholar 

  4. Leuker, C. E., Sonneborn, A., Delbruck, S., and Ernst, J. F. (1997) Sequence and promoter regulation of the PCK1 gene encoding phosphoenolpyruvate carboxykinase of the fungal pathogen Candida albicans, Gene192, 235–240.

    Article  CAS  Google Scholar 

  5. Nobile, C. J., Andes, D. R., Nett, J. E., Smith, F. J., Yue, F., Phan, Q. T., Edwards, J. E., Filler, S. G., and Mitchell, A. P. (2006) Critical role of Bcr1-dependent adhesins in C. albicans biofilm formation in vitro and in vivo, PLoS Pathog.2, e63.

    Article  Google Scholar 

  6. Nobile, C. J., Solis, N., Myers, C. L., Fay, A. J., Deneault, J. S., Nantel, A., Mitchell, A. P., and Filler, S. G. (2008) Candida albicans transcription factor Rim101 mediates pathogenic interactions through cell wall functions, Cell Microbiol10, 2180–2196.

    Article  CAS  Google Scholar 

  7. Nobile, C. J., Nett, J. E., Hernday, A. D., Homann, O. R., Deneault, J. S., Nantel, A., Andes, D. R., Johnson, A. D., and Mitchell, A. P. (2009) Biofilm matrix regulation by Candida albicans Zap1, PLoS biology7, e1000133.

    Article  Google Scholar 

  8. Park, Y. N., and Morschhauser, J. (2005) Tetracycline-inducible gene expression and gene deletion in Candida albicans, Eukaryot Cell4, 1328–1342.

    Article  CAS  Google Scholar 

  9. Fu, Y., Luo, G., Spellberg, B. J., Edwards, J. E., Jr., and Ibrahim, A. S. (2008) Gene overexpression/suppression analysis of candidate virulence factors of Candida albicans, Eukaryot Cell7, 483–492.

    Article  CAS  Google Scholar 

  10. Saville, S. P., Lazzell, A. L., Monteagudo, C., and Lopez-Ribot, J. L. (2003) Engineered control of cell morphology in vivo reveals distinct roles for yeast and filamentous forms of Candida albicans during infection, Eukaryot. Cell2, 1053–1060.

    CAS  Google Scholar 

  11. Saville, S. P., Thomas, D. P., and Lopez Ribot, J. L. (2006) A role for Efg1p in Candida albicans interactions with extracellular matrices, FEMS Microbiol Lett256, 151–158.

    Article  CAS  Google Scholar 

  12. Uppuluri, P., Pierce, C. G., Thomas, D. P., Bubeck, S. S., Saville, S. P., and Lopez-Ribot, J. L. (2010) The transcriptional regulator Nrg1p controls Candida albicans biofilm formation and dispersion, Eukaryot Cell9, 1531–1537.

    Article  CAS  Google Scholar 

  13. Carlisle, P. L., Banerjee, M., Lazzell, A., Monteagudo, C., Lopez-Ribot, J. L., and Kadosh, D. (2009) Expression levels of a filament-specific transcriptional regulator are sufficient to determine Candida albicans morphology and virulence, Proc Natl Acad Sci USA106, 599–604.

    Article  CAS  Google Scholar 

  14. Sahni, N., Yi, S., Daniels, K. J., Huang, G., Srikantha, T., and Soll, D. R. (2010) Tec1 mediates the pheromone response of the white phenotype of Candida albicans: insights into the evolution of new signal transduction pathways, PLoS biology8, e1000363.

    Article  Google Scholar 

  15. Murad, A. M., Lee, P. R., Broadbent, I. D., Barelle, C. J., and Brown, A. J. (2000) CIp10, an efficient and convenient integrating vector for Candida albicans, Yeast16, 325–327.

    Article  CAS  Google Scholar 

  16. Rigaut, G., Shevchenko, A., Rutz, B., Wilm, M., Mann, M., and Seraphin, B. (1999) A generic protein purification method for protein complex characterization and proteome exploration, Nat Biotechnol17, 1030–1032.

    Article  CAS  Google Scholar 

  17. Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular cloning: a laboratory manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.

    Google Scholar 

  18. Gillum, A. M., Tsay, E. Y., and Kirsch, D. R. (1984) Isolation of the Candida albicans gene for orotidine-5’-phosphate decarboxylase by complementation of S. cerevisiae ura3 and E. coli pyrF mutations, Mol. Gen. Genet.198, 179–182.

    Article  CAS  Google Scholar 

  19. Firon, A., Aubert, S., Iraqui, I., Guadagnini, S., Goyard, S., Prevost, M. C., Janbon, G., and d’Enfert, C. (2007) The SUN41 and SUN42 genes are essential for cell separation in Candida albicans, Mol. Microbiol.66, 1256–1275.

    Google Scholar 

  20. Taylor, R. G., Walker, D. C., and McInnes, R. R. (1993) E. coli host strains significantly affect the quality of small scale plasmid DNA preparations used for sequencing, Nucleic Acids Res21, 1677–1678.

    Article  CAS  Google Scholar 

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Acknowledgements

Work in the laboratories of CdE and CM is supported by the Wellcome Trust (The C. albicans ORFeome project, WT088858MA). Work in the CdE laboratory is also supported by the Agence Nationale de la Recherche (KANJI, ANR-08-MIE-033-01) and the European Commission (EURESFUN, LSHM-CT-2005-518199; FINSysB, PITN-GA-2008-214004). Audrey Nesseir is the recipient of a PhD fellowship of the DIM-Maladies Infectieuses. Vitor Cabral is the recipient of a PhD fellowship of the European Commission (FINSysB, PITN-GA-2008-214004) and Mélanie Legrand and Arnaud Firon were recipients of post-doctoral fellowships of Institut Pasteur (Bourse Roux). We are grateful to Pilar Gutierrez-Escribano for suggesting the microwave procedure for colony PCR.

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Authors and Affiliations

  1. Département Génomes et Génétique, Institut Pasteur, Unité Biologie et Pathogénicité Fongiques, Paris, France

    Vitor Cabral, Murielle Chauvel, Arnaud Firon, Mélanie Legrand, Audrey Nesseir, Sophie Bachellier-Bassi & Christophe d’Enfert

  2. INRA USC2019, Paris, France

    Vitor Cabral, Murielle Chauvel, Arnaud Firon, Mélanie Legrand, Audrey Nesseir, Sophie Bachellier-Bassi & Christophe d’Enfert

  3. School of Medical Sciences, University of Aberdeen, Aberdeen, UK

    Yogesh Chaudhari & Carol A. Munro

Authors
  1. Vitor Cabral
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  2. Murielle Chauvel
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  3. Arnaud Firon
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  4. Mélanie Legrand
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  5. Audrey Nesseir
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  6. Sophie Bachellier-Bassi
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  7. Yogesh Chaudhari
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  8. Carol A. Munro
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  9. Christophe d’Enfert
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Corresponding author

Correspondence to Carol A. Munro .

Editor information

Editors and Affiliations

  1. Institute of Medical Sciences, School of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen, AB25 2ZD, United Kingdom

    Alexandra C. Brand

  2. Institute of Medical Sciences, School of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen, AB25 2ZD, United Kingdom

    Donna M. MacCallum

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Cabral, V. et al. (2012). Modular Gene Over-expression Strategies for Candida albicans . In: Brand, A., MacCallum, D. (eds) Host-Fungus Interactions. Methods in Molecular Biology, vol 845. Humana, Totowa, NJ. https://doi.org/10.1007/978-1-61779-539-8_15

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  • DOI: https://doi.org/10.1007/978-1-61779-539-8_15

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  • Publisher Name: Humana, Totowa, NJ

  • Print ISBN: 978-1-61779-538-1

  • Online ISBN: 978-1-61779-539-8

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