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 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.
Choi
Eun-Sil
ES
Department of Biological Sciences, College of Natural Sciences, Chonnam National University, Gwangju 500-757, Republic of Korea.
Han
Sang-Soo
SS
Cheong
Dea-Eun
DE
Park
Mi-Young
MY
Kim
Jeong-Sun
JS
Kim
Geun-Joong
GJ
D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D008164:Luminescent Proteins / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification; D016296:Mutagenesis; D015202:Protein Engineering; D055550:Protein Stability; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D013050:Spectrometry, Fluorescence; D000097573:Red Fluorescent Protein
D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D000097573:Red Fluorescent Protein