Monomeric red fluorescent protein variants used for imaging studies in different species |
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| Authors: | Müller-Taubenberger Annette Vos Michel J Böttger Angelika Lasi Margherita Lai Frank P L Fischer Markus Rottner Klemens |
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| Affiliation: | 1. Institut für Zellbiologie (ABI), Ludwig-Maximilians-Universität München, Schillerstr. 42, D-80336 München, Germany;2. Department of Cell Biology, Section of Radiation and Stress Cell Biology, UMCG, University of Groningen, Groningen, The Netherlands;3. Institut für Biologie II, Ludwig-Maximilians-Universität München, Großhaderner Str. 2, D-82152 Planegg-Martinsried, Germany;4. Cytoskeleton Dynamics Group, German Research Centre for Biotechnology (GBF), Mascheroder Weg 1, D-38124 Braunschweig, Germany;5. Lehrstuhl für Organische Chemie und Biochemie, Technische Universität München, Lichtenbergstr. 4, D-85747 Garching, Germany;1. Ludwig-Maximilians-Universitaet Muenchen, Institut fuer Zellbiologie (ABI), Schillerstr. 42, D-80336 Muenchen, Germany;2. University of Turin, Department of Clinical and Biological Sciences, I-10043 Orbassano, Italy;1. Department of Forensic Medicine, Ga-Rankuwa Forensic Pathology Service, Sefako Makgatho Health Sciences University, Ga-Rankuwa, PO Box 127, Medunsa, 0208, South Africa;2. Department of Forensic Medicine, Pretoria Forensic Pathology Service, University of Pretoria, Private Bag X323, Pretoria, 0007, South Africa;1. Tokyo University of Pharmacy and Life Sciences, Horinouchi 1432-1, Hachioji 192-0392, Tokyo, Japan;2. Asahina Shinryoujo, Nyufune 314-5, Okabecho, Fujieda, Shizuoka 421-1115, Japan;3. Department of Internal Medicine, Fujieda Municipal General Hospital, Surugadai 4-1-11, Fujieda, Shizuoka 426-8677, Japan;4. Department of Gynecology and Obstetrics, Fujieda Municipal General Hospital, Surugadai 4-1-11, Fujieda, Shizuoka 426-8677, Japan;5. Department of Pediatrics, Fujieda Municipal General Hospital, Surugadai 4-1-11, Fujieda, Shizuoka 426-8677, Japan;3. From the Institute of Molecular Medicine and Genetics, Department of Neurology, Medical College of Georgia, Georgia Regents University, Augusta, Georgia 30912,;4. the Systems Biology Center, National Heart, Lung and Blood Institute and;5. Laboratory of Biochemistry and Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892;1. Department of Legal Medicine, Nippon Medical School, Tokyo, Japan;2. Tsukuba Medical Examiner’s Office, Tsukuba, Japan;3. Department of Radiology, Seirei Fuji Hospital, Fuji, Japan;4. Department of Radiological Technology, Tsukuba Medical Center, Tsukuba, Japan;1. Institute of Cell Dynamics and Imaging and Cells-in-Motion Cluster of Excellence (EXC 1003 — CiM), University of Muenster, Von Esmarchstrasse 56, 48149 Muenster, Germany;1. Department of Internal Medicine, National Taiwan University Hospital, 7 Chung-Shan South Road, Taipei, Taiwan;2. Division of Endocrinology, Taipei Tzu Chi General Hospital, 289 Jianguo Road, Xindian, Taiwan;3. Division of Nephrology, Taipei Tzu Chi General Hospital, 289 Jianguo Road, Xindian, Taiwan;4. Division of Nephrology, Department of Internal Medicine, Taipei Medical University Hospital, 250 Wu-Hsing Street, Taipei, Taiwan;5. Department of Internal Medicine and Medical Research Center, Cardinal Tien Hospital, 362 Zhongzheng Road, Xindian, Taiwan;6. Department of Internal Medicine, National Taiwan University Hospital, Hsin-Chu Branch, 25 Jingguo Road, Hsinchu, Taiwan;7. Department of Internal Medicine, Taipei City Hospital, Zhongxing Branch, 145 Zhengzhou Road, Taipei, Taiwan;8. Department of Internal Medicine, Postal Hospital, 14 Fu-Chuo Street, Taipei, Taiwan;9. Department of Urology, National Taiwan University Hospital, 7 Chung-Shan South Road, Taipei, Taiwan;10. Institute of Population Health Sciences, National Health Research Institutes, 35 Keyan Road, Zhunan, Taiwan;11. TAIPAI, the Taiwan Primary Aldosteronism Investigation Group |
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| Abstract: | 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 beta-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. |
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| Keywords: | Dictyostelium Drosophila Hydra Actin cytoskeleton Fluorescent protein Monomeric RFP B16-F1 |
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