Characterization of Fluorescent Proteins for Three- and Four-Color Live-Cell Imaging in S. cerevisiae

PLoS One. 2016 Jan 4;11(1):e0146120. doi: 10.1371/journal.pone.0146120. eCollection 2016.

Abstract

Saccharomyces cerevisiae are widely used for imaging fluorescently tagged protein fusions. Fluorescent proteins can easily be inserted into yeast genes at their chromosomal locus, by homologous recombination, for expression of tagged proteins at endogenous levels. This is especially useful for incorporation of multiple fluorescent protein fusions into a single strain, which can be challenging in organisms where genetic manipulation is more complex. However, the availability of optimal fluorescent protein combinations for 3-color imaging is limited. Here, we have characterized a combination of fluorescent proteins, mTFP1/mCitrine/mCherry for multicolor live cell imaging in S. cerevisiae. This combination can be used with conventional blue dyes, such as DAPI, for potential four-color live cell imaging.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / analysis*
  • Bacterial Proteins / genetics
  • Coloring Agents / analysis
  • DNA, Mitochondrial / analysis
  • Fluorescent Dyes / analysis*
  • Genes, Reporter
  • Genetic Vectors
  • Green Fluorescent Proteins / analysis*
  • Green Fluorescent Proteins / genetics
  • Indoles / analysis
  • Luminescent Proteins / analysis*
  • Luminescent Proteins / genetics
  • Optical Imaging / methods*
  • Plasmids
  • Recombinant Proteins / analysis
  • Saccharomyces cerevisiae / growth & development
  • Saccharomyces cerevisiae / ultrastructure*
  • Single-Cell Analysis / methods*
  • Subcellular Fractions / chemistry

Substances

  • Bacterial Proteins
  • Coloring Agents
  • Cyan Fluorescent Protein
  • DNA, Mitochondrial
  • Fluorescent Dyes
  • Indoles
  • Luminescent Proteins
  • Recombinant Proteins
  • citrine protein, bacteria
  • red fluorescent protein
  • Green Fluorescent Proteins
  • DAPI