Far-red fluorescent tag for protein labelling.
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.
D000199:Actins / Q000737:chemistry; D002478:Cells, Cultured; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry; D005347:Fibroblasts / Q000737:chemistry / Q000166:cytology*; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins; D008856:Microscopy, Fluorescence
D000199:Actins; D002868:Chromosomal Proteins, Non-Histone; D005456:Fluorescent Dyes; D008164:Luminescent Proteins; C047458:fibrillarin; D049452:Green Fluorescent Proteins