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A tandem green-red heterodimeric fluorescent protein with high FRET efficiency
Wiens, M.D.; Shen, Y.; Li, X.; Salem, M.A.; Smisdom, N.; Zhang, W.; Brown, A.; Campbell, R.E. (2016). A tandem green-red heterodimeric fluorescent protein with high FRET efficiency. ChemBioChem 17(24): 2361-2367. https://dx.doi.org/10.1002/cbic.201600492
In: ChemBioChem. Wiley-VCH: Weinheim. ISSN 1439-4227; e-ISSN 1439-7633, meer
Peer reviewed article  

Beschikbaar in  Auteurs 

Author keywords
    dimerization; directed evolution; fluorescence; FRET; proteinengineering

Auteurs  Top 
  • Wiens, M.D.
  • Shen, Y.
  • Li, X.
  • Salem, M.A.
  • Smisdom, N.
  • Zhang, W.
  • Brown, A.
  • Campbell, R.E.

Abstract
    The tetrameric red fluorescent protein from Discosoma sp. coral (DsRed) has previously been engineered to produce dimeric and monomeric fluorescent variants with excitation and emission profiles that span the visible spectrum. The brightest of the effectively monomeric DsRed variants is tdTomato—a tandem fusion of a dimeric DsRed variant. Here we describe the engineering of brighter red (RRvT), green (GGvT), and green–red heterodimeric (GRvT) tdTomato variants. GRvT exhibited 99 % intramolecular FRET efficiency, resulting in long Stokes shift red fluorescence. These new variants could prove useful for multicolor live-cell imaging applications.

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