1. Characterization of Fluorescent Proteins for Three- and Four-Color Live-Cell Imaging in S. cerevisiae.
- Author
-
Higuchi-Sanabria R, Garcia EJ, Tomoiaga D, Munteanu EL, Feinstein P, and Pon LA
- Subjects
- Bacterial Proteins genetics, Coloring Agents analysis, DNA, Mitochondrial analysis, Genes, Reporter, Genetic Vectors, Green Fluorescent Proteins genetics, Indoles analysis, Luminescent Proteins genetics, Plasmids, Recombinant Proteins analysis, Saccharomyces cerevisiae growth & development, Subcellular Fractions chemistry, Red Fluorescent Protein, Bacterial Proteins analysis, Fluorescent Dyes analysis, Green Fluorescent Proteins analysis, Luminescent Proteins analysis, Optical Imaging methods, Saccharomyces cerevisiae ultrastructure, Single-Cell Analysis methods
- 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.
- Published
- 2016
- Full Text
- View/download PDF