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A Tandem Green–Red Heterodimeric Fluorescent Protein with High FRET Efficiency
Author(s) -
Wiens Matthew D.,
Shen Yi,
Li Xi,
Salem M. Alaraby,
Smisdom Nick,
Zhang Wei,
Brown Alex,
Campbell Robert E.
Publication year - 2016
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201600492
Subject(s) - förster resonance energy transfer , fluorescence , green fluorescent protein , protein engineering , tandem , stokes shift , intramolecular force , biophysics , nile red , fluorescent protein , chemistry , fusion protein , biology , recombinant dna , biochemistry , gene , stereochemistry , materials science , physics , composite material , enzyme , quantum mechanics
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.