title: A Tandem Green-Red Heterodimeric Fluorescent Protein with High FRET Efficiency.
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.
authors: Department of Chemistry, University of Alberta, 11227 Saskatchewan Drive, Edmonton, Alberta, T6G 2G2, Canada.
authors: Matthew D
authors: MD
authors: Wiens
authors: Department of Chemistry, University of Alberta, 11227 Saskatchewan Drive, Edmonton, Alberta, T6G 2G2, Canada.
authors: Yi
authors: Y
authors: Shen
authors: Department of Chemistry, University of Alberta, 11227 Saskatchewan Drive, Edmonton, Alberta, T6G 2G2, Canada.
authors: Xi
authors: X
authors: Li
authors: Department of Chemistry, University of Alberta, 11227 Saskatchewan Drive, Edmonton, Alberta, T6G 2G2, Canada.
authors: M Alaraby
authors: MA
authors: Salem
authors: Department of Chemistry, University of Alberta, 11227 Saskatchewan Drive, Edmonton, Alberta, T6G 2G2, Canada.|Present address: Faculty of Medicine and Life Sciences, Universiteit Hasselt, Martelarenlaan 42, 3500, Hasselt, Belgium.
authors: Nick
authors: N
authors: Smisdom
authors: Department of Chemistry, University of Alberta, 11227 Saskatchewan Drive, Edmonton, Alberta, T6G 2G2, Canada.
authors: Wei
authors: W
authors: Zhang
authors: Department of Chemistry, University of Alberta, 11227 Saskatchewan Drive, Edmonton, Alberta, T6G 2G2, Canada.
authors: Alex
authors: A
authors: Brown
authors: Department of Chemistry, University of Alberta, 11227 Saskatchewan Drive, Edmonton, Alberta, T6G 2G2, Canada.
authors: Robert E
authors: http://orcid.org/0000-0003-0604-092X
authors: RE
authors: Campbell
mesh_terms: D000818:Animals; D037421:Anthozoa / Q000378:metabolism; D019281:Dimerization; D031541:Fluorescence Resonance Energy Transfer; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D000097573:Red Fluorescent Protein
chemical_list: D008164:Luminescent Proteins; D011994:Recombinant Proteins; D000097573:Red Fluorescent Protein
keywords: FRET; dimerization; directed evolution; fluorescence; protein engineering