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TitleA Tandem Green-Red Heterodimeric Fluorescent Protein with High FRET Efficiency.
Publication TypeJournal Article
Year of Publication2016
AuthorsWiens, Matthew D., Yi Shen, Xi Li, Alaraby M Salem, Nick Smisdom, Wei Zhang, Alex Brown, and Robert E. Campbell
JournalChembiochem
Volume17
Issue24
Pagination2361-2367
Date Published2016 Dec 14
ISSN1439-7633
KeywordsAnimals, Anthozoa, Dimerization, Fluorescence Resonance Energy Transfer, HeLa Cells, Humans, Luminescent Proteins, Microscopy, Fluorescence, Models, Molecular, Mutagenesis, Site-Directed, Recombinant Proteins
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.

DOI10.1002/cbic.201600492
Alternate JournalChembiochem
PubMed ID27781394