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12. 2020.03.30.015156v1.full.txt - bioRxiv
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Two new coral fluorescent proteins of distinct colors for sharp visualization of cell-cycle progression ======================================================================================================= * Ryoko Ando * Asako Sakaue-Sawano * Keiko Shoda * Atsushi Miyawaki ## Summary We cloned and characterized two new coral fluorescent proteins: h2-3 and 1-41. h2-3 formed an obligate dimeric complex and exhibited bright fluorescence. On the other hand, 1-41 formed a highly multimeric complex and exhibited dim red fluorescence. We engineered 1-41 into AzaleaB5, a practically useful red-emitting fluorescent protein for cellular labeling applications. We fused h2-3 and AzaleaB5 to the ubiquitination domains of human Geminin and Cdt1, respectively, to generate a new color variant of Fucci (Fluorescent Ubiquitination-based Cell-Cycle Indicator): Fucci5. We found Fucci5 provided brighter nuclear labeling for monitoring cell cycle progression than the 1st and 2nd generations that used mAG/mKO2 and mVenus/mCherry, respectively. ## Introduction In recent years, there have been remarkable improvements in our ability to comprehensively unravel the fine details of cellular events. This is owing to the development of the green fluorescent protein from the jellyfish *Aequorea victoria* (avGFP), its spectral variants, such as cyan- and yellow-emitting variants (CFP and YFP, respectively), and GFP-like proteins including red-emitting fluorescent proteins (RFPs) from other organisms....
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