Practical applications of green fluorescent protein ('GFP')-like fluorescent proteins (FPs) from species of the class Anthozoa (sea anemones, corals and sea pens) are strongly restricted owing to their oligomeric nature. Here we suggest a strategy to overcome this problem by the use of two covalently linked identical red FPs as non-oligomerizing fusion tags. We have applied this approach to the dimeric far-red fluorescent protein HcRed1 and have demonstrated superiority of the tandem tag in the in vivo labelling of fine cytoskeletal structures and tiny nucleoli. In addition, a possibility of effective fluorescence resonance energy transfer ('FRET') between enhanced yellow FP mutant ('EYFP') and tandem HcRed1 was demonstrated in a protease assay.
Far-red fluorescent tag for protein labelling
Arkady F. FRADKOV, Vladislav V. VERKHUSHA, Dmitry B. STAROVEROV, Maria E. BULINA, Yurii G. YANUSHEVICH, Vladimir I. MARTYNOV, Sergey LUKYANOV, Konstantin A. LUKYANOV; Far-red fluorescent tag for protein labelling. Biochem J 15 November 2002; 368 (1): 17–21. doi: https://doi.org/10.1042/bj20021191
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