A recyclable Candida albicans URA3 cassette for PCR product-directed gene disruptions

R. Bryce Wilson, Dana Davis, Brian M. Enloe, Aaron P. Mitchell

Research output: Contribution to journalArticle

182 Scopus citations

Abstract

For some time, gene disruptions in Candida albicans have been made with the hisG-URA3-hisG ('Ura-blaster') cassette, which can be re-used in successive transformations of a single strain after homologous excision of URA3. However, the hisG repeats are too large for efficient PCR amplification of the entire cassette, so it cannot be used for PCR product-directed gene disruptions. We describe here a gene disruption cassette, URA3-dpl200, with 200 bp flanking repeats that permit efficient PCR amplification. After transformation and integration to produce both arg5::URA3-dpl200 and rim101::URA3-dpl200 alleles, we find that arg5::dpl200 and rim101::dpl200 segregants, respectively, can be obtained. We have used the cassette to create rim101::dpl200/rim101::URA3-dpl200 mutants exclusively through PCR product-directed disruption. Copyright (C) 2000 John Wiley and Sons, Ltd.

Original languageEnglish (US)
Pages (from-to)65-70
Number of pages6
JournalYeast
Volume16
Issue number1
DOIs
StatePublished - Jan 15 2000

Keywords

  • Candida albicans
  • Gene disruption
  • Ura-blaster

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