title: Characterization of Fluorescent Proteins for Three- and Four-Color Live-Cell Imaging in S. cerevisiae.
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.
authors.affiliation: Department of Pathology and Cell Biology, Columbia University, New York, NY, United States of America.
authors.forename: Ryo
authors.initials: R
authors.lastname: Higuchi-Sanabria
authors.affiliation: Department of Pathology and Cell Biology, Columbia University, New York, NY, United States of America.
authors.forename: Enrique J
authors.initials: EJ
authors.lastname: Garcia
authors.affiliation: 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.
authors.forename: Delia
authors.initials: D
authors.lastname: Tomoiaga
authors.affiliation: Herbert Irving Comprehensive Cancer Center, Columbia University, New York, NY, United States of America.
authors.forename: Emilia L
authors.initials: EL
authors.lastname: Munteanu
authors.affiliation: 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.
authors.forename: Paul
authors.initials: P
authors.lastname: Feinstein
authors.affiliation: Department of Pathology and Cell Biology, Columbia University, New York, NY, United States of America.|Herbert Irving Comprehensive Cancer Center, Columbia University, New York, NY, United States of America.
authors.forename: Liza A
authors.initials: LA
authors.lastname: Pon
mesh_terms: D001426:Bacterial Proteins / Q000032:analysis* / Q000235:genetics; D004396:Coloring Agents / Q000032:analysis; D004272:DNA, Mitochondrial / Q000032:analysis; D005456:Fluorescent Dyes / Q000032:analysis*; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000032:analysis* / Q000235:genetics; D007211:Indoles / Q000032:analysis; D008164:Luminescent Proteins / Q000032:analysis* / Q000235:genetics; D061848:Optical Imaging / Q000379:methods*; D010957:Plasmids; D011994:Recombinant Proteins / Q000032:analysis; D012441:Saccharomyces cerevisiae / Q000254:growth & development / Q000648:ultrastructure*; D059010:Single-Cell Analysis / Q000379:methods*; D013347:Subcellular Fractions / Q000737:chemistry; D000097573:Red Fluorescent Protein
chemical_list: D001426:Bacterial Proteins; D004396:Coloring Agents; D004272:DNA, Mitochondrial; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins; D007211:Indoles; D008164:Luminescent Proteins; D011994:Recombinant Proteins; D000097573:Red Fluorescent Protein; C485182:Cyan Fluorescent Protein; C434479:citrine protein, bacteria; C007293:DAPI