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

Biochem Biophys Res Commun. 2010 Jan 1;391(1):598-603. doi: 10.1016/j.bbrc.2009.11.105. Epub 2009 Nov 20.

Abstract

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.

MeSH terms

  • Escherichia coli / genetics
  • Gene Library
  • Luminescent Proteins / chemistry*
  • Luminescent Proteins / genetics*
  • Luminescent Proteins / isolation & purification
  • Mutagenesis
  • Protein Engineering
  • Protein Stability
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / isolation & purification
  • Red Fluorescent Protein
  • Spectrometry, Fluorescence

Substances

  • Luminescent Proteins
  • Recombinant Fusion Proteins