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Title: One-step targeted gene deletion in Candida albicans haploids
Authors: Zeng, G.
Wang, Y.-M.
Chan, F.Y.
Wang, Y. 
Issue Date: Feb-2014
Citation: Zeng, G., Wang, Y.-M., Chan, F.Y., Wang, Y. (2014-02). One-step targeted gene deletion in Candida albicans haploids. Nature Protocols 9 (2) : 464-473. ScholarBank@NUS Repository.
Abstract: The recent discovery of haploids in Candida albicans and the construction of tool strains carrying multiple auxotrophic markers have enabled, for the first time, performing one-step gene deletions in this fungal human pathogen. This breakthrough promises to greatly facilitate the molecular and genetic study of C. albicans biology and pathogenicity. However, the construction of gene-deletion mutants in C. albicans haploids involves many technical difficulties, particularly low transformation efficiency and autodiploidization. Here we describe a highly effective protocol for designing and performing one-step gene deletion in C. albicans haploids, which takes ∼11 d to complete (not including plasmid construction, which may take ∼2 weeks). A gene deletion cassette is constructed on a plasmid and subsequently released for transformation by lithium acetate incubation or electroporation. Desired gene-deletion mutants are identified and their ploidy is assessed simultaneously by colony PCR before final confirmation by flow cytometry. © 2014 Nature America, Inc. All rights reserved.
Source Title: Nature Protocols
ISSN: 17542189
DOI: 10.1038/nprot.2014.029
Appears in Collections:Staff Publications

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