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A Luciferase Reporter for Gene Expression Studies and Dynamic Imaging of Superficial Candida albicans Infections

  • Donatella Pietrella
  • Brice Enjalbert
  • Ute Zeidler
  • Sadri Znaidi
  • Anna Rachini
  • Anna Vecchiarelli
  • Christophe d’EnfertEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 845)

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.

Key words

Luciferase Gaussia princeps Imaging Reporter Gene fusion Vulvovaginal candidiasis Subcutaneous candidiasis Superficial infection 

Notes

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).

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Donatella Pietrella
    • 1
  • Brice Enjalbert
    • 2
  • Ute Zeidler
    • 3
  • Sadri Znaidi
    • 3
  • Anna Rachini
    • 1
  • Anna Vecchiarelli
    • 1
  • Christophe d’Enfert
    • 4
    • 5
    Email author
  1. 1.Microbiology Section, Department of Experimental Medicine and Biochemical SciencesUniversity of PerugiaPerugiaItaly
  2. 2.Institut National des Sciences AppliquéesToulouseFrance
  3. 3.Institut Pasteur, Unité Biologie et Pathogénicité FongiquesParisFrance
  4. 4.Département Génomes et GénétiqueInstitut Pasteur, Unité Biologie et Pathogénicité FongiquesParisFrance
  5. 5.INRA USC2019ParisFrance

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