Direct transformation of a clinical isolate of Candida parapsilosis using a dominant selection marker

Attila Gácser, Siegfried Salomon, Wilhelm Schäfer

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Candida parapsilosis is a human pathogenic fungus with increasing importance, particularly in nosocomial infections. For detailed molecular genetic explorations of prototrophic clinical isolates of C. parapsilosis, we developed an efficient transformation system based on a dominant selectable marker. The gene encoding resistance to mycophenolic acid (MPA) was used for selection in yeast transformation. C. parapsilosis cells were transformed with a plasmid vector containing the Candida albicans inosine monophosphate dehydrogenase gene (IMH3) responsible for mycophenolic acid resistance. Transformation was carried out both by electroporation and by the lithium acetate (LiAc) method. The LiAc method resulted in very poor transformation efficiency, while the modified electroporation method yielded a high number of mitotically stable transformants exhibiting unambiguous MPA resistance. Two hundred transformants were analysed for the presence of the C. albicans IMH3r gene by polymerase chain reaction. Integration of single or multiple plasmid copies into the genomic DNA of C. parapsilosis was determined by Southern hybridization. To our knowledge, the present study is the first report about a method based on a dominant selectable marker for the transformation of a prototrophic, clinical isolate of C. parapsilosis. The described technique may prove to be an efficient tool for the examination of the biology and virulence of this pathogenic yeast.

Original languageEnglish
Pages (from-to)117-121
Number of pages5
JournalFEMS microbiology letters
Volume245
Issue number1
DOIs
Publication statusPublished - Apr 1 2005

Keywords

  • Candida parapsilosis
  • Human pathogen
  • Mycophenolic acid resistance
  • Pathogenic yeast
  • Transformation system

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology
  • Genetics

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