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GFP-like chromoproteins as a source of far-red fluorescent proteins11 The accession numbers of genes described in this article are: cgCP (AF363775), hcCP (AF363776), cpCP (AF383155), gtCP (AF383156).

Authors
Journal
FEBS Letters
0014-5793
Publisher
Wiley Blackwell (John Wiley & Sons)
Publication Date
Volume
507
Issue
1
Identifiers
DOI: 10.1016/s0014-5793(01)02930-1
Keywords
  • Green Fluorescent Protein
  • Anthozoa
  • Drfp583
  • Dsred
  • Multi-Color Labeling
Disciplines
  • Biology

Abstract

Abstract 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.

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