Yeast-enhanced green fluorescent protein (yEGFP): a reporter of gene expression in Candida albicans.
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.
Cormack
Brendan P
BP
Department of Microbiology and Immunology, Stanford University School of Medicine,Stanford, CA 94305-5402,USA.
Bertram
Gwyneth
G
Molecular and Cell Biology, University of Aberdeen, Institute of Medical Sciences,Foresterhill, Aberdeen AB25 2ZD,UK.
Egerton
Mark
M
Zeneca Pharmaceuticals, Alderley Park, Macclesfield SK10 4TG,UK.
Gow
Neil A R
NAR
Zeneca Pharmaceuticals, Alderley Park, Macclesfield SK10 4TG,UK.
Falkow
Stanley
S
Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases,Hamilton, MT 59840,USA.|Department of Microbiology and Immunology, Stanford University School of Medicine,Stanford, CA 94305-5402,USA.
Brown
Alistair J P
AJP
Molecular and Cell Biology, University of Aberdeen, Institute of Medical Sciences,Foresterhill, Aberdeen AB25 2ZD,UK.
D000818:Animals; D001483:Base Sequence; D002176:Candida albicans / Q000235:genetics*; D002177:Candidiasis / Q000382:microbiology; D003062:Codon; D005069:Evaluation Studies as Topic; D005434:Flow Cytometry; D005453:Fluorescence; D015870:Gene Expression; D017930:Genes, Reporter; D005813:Genes, Synthetic; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013045:Species Specificity
D003062:Codon; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins