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GFP‐like chromoproteins as a source of far‐red fluorescent proteins
Author(s) -
Gurskaya Nadya G.,
Fradkov Arkady F.,
Terskikh Alexey,
Matz Mikhail V.,
Labas Yulii A.,
Martynov Vladimir I.,
Yanushevich Yurii G.,
Lukyanov Konstantin A.,
Lukyanov Sergey A.
Publication year - 2001
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(01)02930-1
Subject(s) - fluorescence , chemistry , green fluorescent protein , biophysics , mutagenesis , mutant , biochemistry , biology , gene , optics , physics
We have employed a new approach to generate novel fluorescent proteins (FPs) from red absorbing chromoproteins. An identical single amino acid substitution converted novel chromoproteins from the species Anthozoa ( Heteractis crispa , Condylactis gigantea , and Goniopora tenuidens ) into far‐red FPs (emission λ max =615–640 nm). Moreover, coupled site‐directed and random mutagenesis of the chromoprotein from H. crispa resulted in a unique far‐red FP (HcRed) that exhibited bright emission at 645 nm. A clear red shift in fluorescence of HcRed, compared to drFP583 (by more than 60 nm), makes it an ideal additional color for multi‐color labeling. Importantly, HcRed is excitable by 600 nm dye laser, thus promoting new detection channels for multi‐color flow cytometry applications. In addition, we generated a dimeric mutant with similar maturation and spectral properties to tetrameric HcRed.