Protein mobility in the cytoplasm of Escherichia coli
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...The diffusion rates of macromolecules inside a cell are generally slow and limitin...
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Cites background from "Protein mobility in the cytoplasm o..."
...ense they complement each other. Combining FRAP and a photoactivation technique allowed for a non-invasive measurement of the motion of endogenously expressed GFP in the cytoplasm of E. coli bacteria [250]. The apparent diffusion constant was obtained as 7:7 ±2:5µm2/s at the time scale of some 0.1s, which is about 11 times smaller than for free diffusion in water and signifi- cantly lower than in eukary...
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"Protein mobility in the cytoplasm o..." refers methods in this paper
...One of the most useful techniques for studying cytoplasmic diffusion in eukaryotic cells and cell membranes has been the method of “fluorescence recovery after photobleaching” (FRAP) (3, 16)....
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...These bacteria are smaller than the eukaryotic cells previously studied by FRAP, and it is difficult to introduce fluorescently labeled molecules into them....
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...Use of the FRAP technique with image data in the small quasicylindrical geometry of the bacterial cell called for a method of analysis which considers the concentration of GFP throughout the cell rather than only in the photobleached region (see Materials and Methods)....
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...Like fluorochromes used in previous FRAP experiments, GFP can be irreversibly photobleached with sufficiently intense illumination (25)....
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...FRAP experiments have difficulty distinguishing between normal diffusion with a fixed “immobile fraction” and subdiffusion (11)....
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Additional excerpts
...In addition, the following E. coli strains transformed with pMGS053 were examined: AB1157 (9), M15(pREP4) (Qiagen), MC1000 (5), MC1061 (5), MG1655 (13), and RP437 (18)....
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...were examined: AB1157 (9), M15(pREP4) (Qiagen), MC1000 (5), MC1061 (5),...
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