Monomeric red fluorescent protein variants used for imaging studies in different species

• Published in: European journal of cell biology Vol. 85; no. 9; pp. 1119 - 1129
• Main Authors: Müller-Taubenberger, Annette, Vos, Michel J., Böttger, Angelika, Lasi, Margherita, Lai, Frank P.L., Fischer, Markus, Rottner, Klemens
• Format: Journal Article
• Language: English
• Published: Germany Elsevier GmbH 01.09.2006
• Subjects: Actin cytoskeleton; Animals; B16-F1; Cell Culture Techniques; Cell Line; Dictyostelium; Dictyostelium - cytology; Dictyostelium - genetics; Dictyostelium - metabolism; Drosophila; Drosophila melanogaster; Fluorescent protein; Humans; Hydra; Hydra - cytology; Hydra - genetics; Hydra - metabolism; Luminescent Proteins - genetics; Luminescent Proteins - metabolism; Mice; Microscopy, Fluorescence; Monomeric RFP; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Red Fluorescent Protein; Monomeric RFP; Dictyostelium; Fluorescent protein; Drosophila; B16-F1; Actin cytoskeleton; Hydra
• ISSN: 0171-9335; 1618-1298
• DOI: 10.1016/j.ejcb.2006.05.006

Fluorescent proteins have proven to be excellent tools for live-cell imaging studies. In addition to green fluorescent protein (GFP) and its variants, recent progress was achieved in the development of monomeric red fluorescent proteins (mRFPs) that show improved properties in respect to maturation and intracellular fluorescence. mRFPmars, a red fluorescent protein designed especially for the use in Dictyostelium, has been employed to tag different proteins for live-cell investigations in Dictyostelium. mRFPruby, which differs in sequence from mRFPmars in four amino acids, has a codon usage optimised for the application in mammalian cells. Here, we show that both mRFP variants can also be applied for localisation studies in other organisms. mRFPmars was expressed in Hydra and fused to the Bcl-2 family protein Bax. mRFPruby in combination with histone 2B was expressed in Drosophila S2 cells to monitor mitosis. Using mouse cell lines, mRFPruby fused to β-actin was assayed with high spatial resolution to study details of actin cytoskeleton dynamics. In addition, we demonstrate that both mRFP variants are also suitable for dual-colour microscopy in the different species.