Yeast-enhanced green fluorescent protein (yEGFP): A reporter of gene expression in Candida albicans

Brendan P. Cormack*, Gwyneth Bertram, Mark Egerton, Neil A.R. Gow, Stanley Falkow, Alistair J.P. Brown

*Corresponding author for this work

Research output: Contribution to journalArticle

468 Citations (Scopus)

Abstract

The green fluorescent protein (GFP) of Aequorea victoria has been developed here as a reporter for gene expression and protein localization in Candida albicans. When wild-type (wt) GFP was expressed in C. albicans, it was not possible to detect fluorescence or a translation product for the wt protein. Since this was probably due in part to the presence of the non-canonical CTG serine codon in the Aequorea sequence, this codon was changed to the leucine codon TTG. C. albicans cells expressing this construct contained CFP mRNA but were non-fluorescent and contained no detectable translation product. Hence a codon-optimized GFP gene was constructed in which all of the 239 amino acids are encoded by optimal codons for C. albicans. In this gene were also incorporated two previously identified mutations in the chromophore that increase GFP fluorescence. C. albicans cells expressing this yeast-enhanced GFP gene (yEGFP3) are fluorescent and contain CFP protein. yEGFP3 can be used as a versatile reporter of gene expression in C. albicans and Saccharomyces cerevisiae and the optimized GFP described here should have broad applications in these and other fungal species.

Original languageEnglish
Pages (from-to)303-311
Number of pages9
JournalMicrobiology
Volume143
Issue number2
DOIs
Publication statusPublished - 1 Feb 1997

Keywords

  • Candida albicans
  • Enhanced GFP
  • Green fluorescent protein
  • Saccharomyces cerevisiae

ASJC Scopus subject areas

  • Microbiology

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