Targeted gene deletion in Candida albicans wild-type strains by MPAR flipping.

Joachim Morschhäuser, Peter Staib, Gerwald Koehler

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Many genetic and phenotypic changes occur during the development of drug resistance in fungi. A straightforward approach to assess the contribution of a specific gene to drug resistance is to examine its inactivation in a resistant isolate and to analyze the effect of the mutation on the resistance phenotype. The generation of knockout mutants in the diploid yeast Candida albicans requires two rounds of gene replacement to inactivate both alleles of a target gene. Because auxotrophic markers are not useful for the genetic manipulation of wild-type, clinical isolates, dominant selection markers are required. In this chapter, we describe the MPAR flipping method that combines dominant selection with recombinase-mediated marker recycling for targeted inactivation of specific genes in C. albicans wild-type strains. Using the MPAR flipper makes drug-resistant clinical isolates amenable to genetic manipulation, a prerequisite for the study of causal relationships between specific genes and drug resistance.

Original languageEnglish
Pages (from-to)35-44
Number of pages10
JournalMethods in molecular medicine
Volume118
StatePublished - 1 Jan 2005

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Gene Deletion
Candida albicans
Drug Resistance
Genes
Recombinases
Recycling
Gene Silencing
Diploidy
Fungi
Yeasts
Alleles
Phenotype
Mutation
Pharmaceutical Preparations

Cite this

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Targeted gene deletion in Candida albicans wild-type strains by MPAR flipping. / Morschhäuser, Joachim; Staib, Peter; Koehler, Gerwald.

In: Methods in molecular medicine, Vol. 118, 01.01.2005, p. 35-44.

Research output: Contribution to journalArticle

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