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A brilliant monomeric red fluorescent protein to visualize cytoskeleton dynamics in Dictyostelium
Author(s) -
Fischer Markus,
Haase Ilka,
Simmeth Evelyn,
Gerisch Günther,
Müller-Taubenberger Annette
Publication year - 2004
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/j.febslet.2004.09.084
Subject(s) - green fluorescent protein , fluorescence , cytoskeleton , fluorescence microscope , dictyostelium discoideum , biophysics , microbiology and biotechnology , microtubule , chemistry , dictyostelium , biology , biochemistry , cell , physics , quantum mechanics , gene
Red fluorescent proteins (RFPs) combined with GFP are attractive probes for double‐fluorescence labeling of proteins in live cells. However, the application of these proteins is restrained by stable oligomer formation and by their weak fluorescence in vivo. Previous attempts to eliminate these problems by mutagenesis of RFP from Discosoma (DsRed) resulted in the monomeric mRFP1 and in the tetrameric RedStar RFP, which is distinguished by its enhanced fluorescence in vivo. Based on these mutations, we have generated an enhanced monomeric RFP, mRFPmars, and report its spectral properties. Together with green fluorescent labels, we used mRFPmars to visualize filamentous actin structures and microtubules in Dictyostelium cells. This enhanced RFP proved to be suitable to monitor the dynamics of cytoskeletal proteins in cell motility, mitosis, and endocytosis using dual‐wavelength fluorescence microscopy.