A molecular genetic system for the pathogenic yeast Candida dubliniensis.
Candida dubliniensis is a recently described pathogenic yeast of the genus Candida that is closely related to Candida albicans but differs from it in several phenotypic and genotypic characteristics, including putative virulence traits, which may explain differences in the spectrum of diseases caused by the two species. In contrast to C. albicans, a molecular genetic system to study virulence of C. dubliniensis is lacking. We have developed a system for the genetic transformation of C. dubliniensis that is based on the use of the dominant selection marker MPA(R) from C. albicans that confers resistance to mycophenolic acid (MPA). Using this transformation system, a GFP (green fluorescent protein) reporter gene that was genetically engineered for functional expression in C. albicans and placed under control of the inducible C. albicans SAP2 (secreted aspartic proteinase) promoter was integrated into the C. dubliniensis genome. MPA-resistant transformants containing the SAP2P-GFP fusion fluoresced under SAP2-inducing conditions but not under SAP2-repressing conditions. These results demonstrate that the MPA(R) selection marker is useful for transformation of C. dubliniensis wild-type strains, that the GFP reporter gene is functionally expressed in C. dubliniensis, and that the C. albicans SAP2 promoter can be used for controlled gene expression in C. dubliniensis. These genetic tools will allow the dissection of the differences in virulence characteristics between the two pathogenic yeast species at the molecular level.
Staib
P
P
Zentrum für Infektionsforschung, Universität Würzburg, Germany.
Michel
S
S
Köhler
G
G
Morschhäuser
J
J
D016282:Aspartic Acid Endopeptidases / Q000235:genetics; D002175:Candida / Q000187:drug effects / Q000235:genetics* / Q000472:pathogenicity; D002176:Candida albicans / Q000235:genetics; D003470:Culture Media / Q000494:pharmacology; D004352:Drug Resistance, Microbial / Q000235:genetics; D005656:Fungal Proteins; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D009173:Mycophenolic Acid / Q000494:pharmacology; D011401:Promoter Regions, Genetic / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D014170:Transformation, Genetic; D014774:Virulence / Q000235:genetics
D003470:Culture Media; D005656:Fungal Proteins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D016282:Aspartic Acid Endopeptidases; C090753:SAP2 protein, Candida; D009173:Mycophenolic Acid