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

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

Author information

1
Department of Pathology and Cell Biology, Columbia University, New York, NY, United States of America.
2
Department of Biological Sciences, Hunter College and The Graduate Center Biochemistry, Biology and Biopsychology and Behavioral Neuroscience Programs, CUNY, New York, NY 10065, United States of America.
3
Herbert Irving Comprehensive Cancer Center, Columbia University, New York, NY, United States of America.

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.

PMID:
26727004
PMCID:
PMC4699809
DOI:
10.1371/journal.pone.0146120
[Indexed for MEDLINE]
Free PMC Article

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