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Schizosaccharomyces pombe  is an outstanding model organism for cell biological investigations, yet the range of useful and well-characterized fluorescent proteins (XFPs) is limited. We generated and characterized three recoded fluorescent proteins for 3-color analysis in  S . pombe , Super-folder GFP, monomeric Kusabira Orange 2 and E2Crimson. Upon optimization and expression in  S .  pombe , the three proteins enabled sensitive simultaneous 3-color detection capability. Furthermore, we describe a strategy that combines a pulse-chase approach and mathematical modeling to quantify the maturation kinetics of these proteins  in vivo . We observed maturation kinetics in  S .  pombe  that are expected from those described for these proteins  in vitro  and/or in other cell types, but also unpredicted behaviors. Our studies provide a kinetically-characterized, integrated three-color XFP toolbox for  S .  pombe .

plos one

Sensitive and Quantitative Three-Color Protein Imaging in Fission Yeast Using Spectrally Diverse, Recoded Fluorescent Proteins with Experimentally-Characterized In Vivo Maturation Kinetics