mKikGR,a Monomeric Photoswitchable Fluorescent Protein |
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| Authors: | Satoshi Habuchi Hidekazu Tsutsui Anna B Kochaniak Atsushi Miyawaki Antoine M van Oijen |
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| Institution: | 1. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts, United States of America.; 2. Laboratory for Cell Function Dynamics, Brain Science Institute, RIKEN, Wako, Saitama, Japan.; 3. Graduate School of Science and Engineering, Tokyo Institute of Technology, Tokyo, Japan.; 4. Graduate Program in Biophysics, Harvard University, Cambridge, Massachusetts, United States of America.;Massachusetts Institute of Technology, United States of America |
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| Abstract: | The recent demonstration and utilization of fluorescent proteins whose fluorescence can be switched on and off has greatly expanded the toolkit of molecular and cell biology. These photoswitchable proteins have facilitated the characterization of specifically tagged molecular species in the cell and have enabled fluorescence imaging of intracellular structures with a resolution far below the classical diffraction limit of light. Applications are limited, however, by the fast photobleaching, slow photoswitching, and oligomerization typical for photoswitchable proteins currently available. Here, we report the molecular cloning and spectroscopic characterization of mKikGR, a monomeric version of the previously reported KikGR that displays high photostability and switching rates. Furthermore, we present single-molecule imaging experiments that demonstrate that individual mKikGR proteins can be localized with a precision of better than 10 nanometers, suggesting their suitability for super-resolution imaging. |
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