Generation of a fast maturating red fluorescent protein by a combined approach of elongation mutagenesis and functional salvage screening

Fluorescent proteins that can be useful as indicators or reporters must have rapid maturation time, high quantum yield and photobleaching stability. A red fluorescent protein DsRed that has a high quantum yield and photostability has an innately slow maturation time when compared to other fluorescen...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 2010-01, Vol.391 (1), p.598-603
Main Authors: Choi, Eun-Sil, Han, Sang-Soo, Cheong, Dea-Eun, Park, Mi-Young, Kim, Jeong-Sun, Kim, Geun-Joong
Format: Article
Language:English
Subjects:
60 APPLIED LIFE SCIENCES
DsRed
ELONGATION
Elongation mutagenesis
Escherichia coli - genetics
FERROMAGNETIC RESONANCE
FLUORESCENCE
FmRed
Gene Library
Luminescent Proteins - chemistry
Luminescent Proteins - genetics
Luminescent Proteins - isolation & purification
MATURATION
Maturation time
MUTAGENESIS
Protein Engineering
Protein Stability
PROTEINS
Recombinant Fusion Proteins - chemistry
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - isolation & purification
Red fluorescence
Red Fluorescent Protein
SCREENING
Spectrometry, Fluorescence
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Summary:Fluorescent proteins that can be useful as indicators or reporters must have rapid maturation time, high quantum yield and photobleaching stability. A red fluorescent protein DsRed that has a high quantum yield and photostability has an innately slow maturation time when compared to other fluorescence proteins. In this study, we combined a functional salvage screen (FSS) and elongation mutagenesis to obtain a DsRed variant that maintained structural features closely linked with a high quantum yield and photostability and evolved to have a rapid maturation time. It is expected that the variant generated here, FmRed (fast maturating red fluorescent protein), will be widely used as an indicator or reporter because it maintained traits superior to that of the wild-type protein and also matured rapidly.
ISSN:0006-291X
1090-2104
DOI:10.1016/j.bbrc.2009.11.105