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Molecular analysis of the poly-telomeric CTA2 gene family of Candida albicans: Elucidation of genetic or epigenetic regulation of their differential expression, localisation and functional analyses in vitro and during the early phase of colonisation on porcine intestinal epithelium
Antragsteller
Professor Dr. Jürgen Wendland
Fachliche Zuordnung
Parasitologie und Biologie der Erreger tropischer Infektionskrankheiten
Förderung
Förderung von 2004 bis 2008
Projektkennung
Deutsche Forschungsgemeinschaft (DFG) - Projektnummer 5426768
C. albicans is an opportunistic fungal pathogen that is frequently found in the normal gastrointestinal flora of humans. In immuno-compromised patients or in patients with an altered gut flora, e.g. due to antibiotic treatment, colonization and invasion by C. albicans may occur. Particularly infection of the blood stream by C. albicans (candidaemia) often causes death. We have established an in vitro test system to study the first steps during colonization of C. albicans on the molecular level. We use porcine intestinal epithelium (PIE) of freshly slaughtered pigs, which is inoculated with preincubated C. albicans strains. Under these conditions wildtype cells produce germ tubes after 3h which adhere to the PIE. Cawal1 mutant cells, which under standard laboratory conditions do not form hyphae, show at least adhesion to the epithelium. Using the DNA-array technology (based on the Eurogentec DNA-arrays) genes that are differentially expressed during these early stages of infection (e.g. as compared to a efg1/cph1 null strain) and thus contribute to adhesion and/or hyphal induction will be identified. These differentially expressed genes will be functionally analyzed using PCR-based methods. This includes the generation of deletion strains as well as the subcellular localization of the proteins via GFP-fusions.
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