Abstract
Real-time imaging of fungal infections is becoming integral to the study of host–pathogen interactions, as it allows monitoring of the spatial and temporal progression of pathogen growth or of the host response in a single animal as well as reducing the number of animals used to obtain significant data. We present different applications of a novel luciferase reporter gene constructed from the coding sequences of the Candida albicans PGA59 gene, encoding a GPI-linked cell wall protein, and the Gaussia princeps luciferase gene. Upon addition of the coelenterazine substrate, light produced by the surface-exposed luciferase can be used to quantify gene expression from a variety of C. albicans promoters as well as monitoring cutaneous, subcutaneous, and vaginal infections.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Hutchens, M., and Luker, G. D. (2007) Applications of bioluminescence imaging to the study of infectious diseases, Cell Microbiol9, 2315–2322.
Doyle, T. C., Nawotka, K. A., Kawahara, C. B., Francis, K. P., and Contag, P. R. (2006) Visualizing fungal infections in living mice using bioluminescent pathogenic Candida albicans strains transformed with the firefly luciferase gene, Microbial pathogenesis40, 82–90.
Doyle, T. C., Nawotka, K. A., Purchio, A. F., Akin, A. R., Francis, K. P., and Contag, P. R. (2006) Expression of firefly luciferase in Candida albicans and its use in the selection of stable transformants, Microbial pathogenesis40, 69–81.
Tannous, B. A., Kim, D. E., Fernandez, J. L., Weissleder, R., and Breakefield, X. O. (2005) Codon-optimized Gaussia luciferase cDNA for mammalian gene expression in culture and in vivo, Mol Ther11, 435–443.
Moreno-Ruiz, E., Ortu, G., de Groot, P. W., Cottier, F., Loussert, C., Prevost, M. C., de Koster, C., Klis, F. M., Goyard, S., and d’Enfert, C. (2009) The GPI-modified proteins Pga59 and Pga62 of Candida albicans are required for cell wall integrity, Microbiology155, 2004–2020.
Enjalbert, B., Rachini, A., Vediyappan, G., Pietrella, D., Spaccapelo, R., Vecchiarelli, A., Brown, A. J., and d’Enfert, C. (2009) A multifunctional, synthetic Gaussia princeps luciferase reporter for live imaging of Candida albicans infections, Infect Immun77, 4847–4858.
Pietrella, D., Rachini, A., Torosantucci, A., Chiani, P., Brown, A. J., Bistoni, F., Costantino, P., Mosci, P., d’Enfert, C., Rappuoli, R., Cassone, A., and Vecchiarelli, A. (2010) A beta-glucan-conjugate vaccine and anti-beta-glucan antibodies are effective against murine vaginal candidiasis as assessed by a novel in vivo imaging technique, Vaccine28, 1717–1725.
d’Enfert, C., Vecchiarelli, A., and Brown, A. J. (2010) Bioluminescent fungi for real-time monitoring of fungal infections, Virulence1, 174–176.
Vediyappan, G., Rossignol, T., and d’Enfert, C. (2010) Interaction of Candida albicans biofilms with antifungals: transcriptional response and binding of antifungals to beta-glucans, Antimicrob Agents Chemother54, 2096–2111.
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.
Acknowledgements
We are grateful to Al Brown for his contributions to the development of the luciferase reporter. Work in the laboratories of CdE and AV is supported by the European Commission (Galar Fungail 2 Marie Curie Research Training Network, MRTN-CT-2003-504148; FINSysB Marie Curie Initial Training Network, PITN-GA-2008-214004). Work in the laboratory of CdE is also supported by the Agence Nationale de la Recherche (KANJI, ANR-08-MIE-033-01). Brice Enjalbert was the recipient of a postdoctoral fellowship of the European Commission (Galar Fungail 2, MRTN-CT-2003-504148). UZ is the recipient of a postdoctoral fellowship of the program Carnot Maladies Infectieuses. SZ is the recipient of a postdoctoral fellowship of the European Commission (FINSysB, PITN-GA-2008-214004).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Pietrella, D. et al. (2012). A Luciferase Reporter for Gene Expression Studies and Dynamic Imaging of Superficial Candida albicans Infections. 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_39
Download citation
DOI: https://doi.org/10.1007/978-1-61779-539-8_39
Published:
Publisher Name: Humana, Totowa, NJ
Print ISBN: 978-1-61779-538-1
Online ISBN: 978-1-61779-539-8
eBook Packages: Springer Protocols