Expression and analysis of the green fluorescent protein gene in the fission yeast Schizosaccharomyces pombe.
This report demonstrates that the Aequorea victoria green fluorescence protein (gfp) gene product will fluoresce in the fission yeast Schizosaccharomyces pombe when expressed from an episomal expression vector. Fluorescence was readily detectable at both the colony and single cell level. Application of fluorescence-activated cell sorting (FACS) techniques showed that gfp-expressing cells could be detected when they were as rare as 1% of a total yeast population. Quantitative analysis of gfp-expressing cells constituting as little as 5% of a total population was possible. These observations establish the suitability of the gfp gene for use in S. pombe and, in combination with FACS, offers an experimental strategy for quantitative analysis of gene expression in yeast populations.
Atkins
D
D
R. W. Johnson Pharmaceutical Research Institute, Sydney NSW, Australia.
Izant
J G
JG
D001483:Base Sequence; D002469:Cell Separation / Q000379:methods*; D003001:Cloning, Molecular; D005434:Flow Cytometry / Q000379:methods; D005453:Fluorescence; D005656:Fungal Proteins / Q000235:genetics; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D012568:Schizosaccharomyces / Q000235:genetics* / Q000378:metabolism; D029702:Schizosaccharomyces pombe Proteins; D014170:Transformation, Genetic
D005656:Fungal Proteins; D008164:Luminescent Proteins; C093953:NMT1 protein, S pombe; D011994:Recombinant Proteins; D029702:Schizosaccharomyces pombe Proteins; D049452:Green Fluorescent Proteins