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

Microbiology. 1997 Feb;143 ( Pt 2):303-11. doi: 10.1099/00221287-143-2-303.

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 GFP 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 GFP 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.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Candida albicans / genetics*
  • Candidiasis / microbiology
  • Codon
  • Evaluation Studies as Topic
  • Flow Cytometry
  • Fluorescence
  • Gene Expression
  • Genes, Reporter*
  • Genes, Synthetic
  • Green Fluorescent Proteins
  • Luminescent Proteins / biosynthesis*
  • Luminescent Proteins / genetics
  • Mice
  • Molecular Sequence Data
  • Saccharomyces cerevisiae / genetics
  • Species Specificity

Substances

  • Codon
  • Luminescent Proteins
  • Green Fluorescent Proteins

Associated data

  • GENBANK/U73901