Springer Protocols: Abstract: Localization and Mobility of Bacterial Proteins by Confocal Microscopy and Fluorescence Recovery After Photobleaching

Localization and Mobility of Bacterial Proteins by Confocal Microscopy and Fluorescence Recovery After Photobleaching

By: Conrad W. Mullineaux²

Abstract

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This chapter describes the use of laser-scanning confocal fluorescence microscopy for determining the localization of fluorescently tagged proteins within bacterial cells, discussing the problems caused by the limited resolution of an optical microscope. It also explains a relatively simple method for using fluorescence recovery after photobleaching (FRAP) to observe and quantify the diffusion of fluorescently tagged proteins in bacterial cells. The techniques are illustrated with reference to measurements on green fluorescent protein (GFP)-tagged proteins in Escherichia coli.

Affiliation(s): (2) School of Biological and Chemical Sciences, Queen Mary, University of London, London, UK

Book Title: Protein Targeting Protocols

Series: Methods in Molecular Biology | Volume: 390 | Pub. Date: May-21-2007 | Page Range: 3-16 | DOI: 10.1007/978-1-59745-466-7_1

Subject: Protein Science

Key Words: Bacterium - confocal microscopy - cytoplasm - diffusion - Escherichia coli - fluorescence recovery after photobleaching - green fluorescent protein - periplasm - plasma membrane

References

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