One of the most important recent discoveries in science is the green fluorescent protein, isolated from a bioluminescent jellyfish. The gene that codes for this protein is active in any type of cell, from microbes to humans, producing a "day-glo" dye visible as a bright green light in the microscope. The power of the gene is that by using recombinant DNA technology. GFP can be spliced to any protein, essentially painting that protein green. When such a spliced protein is inside a cell. Further, time-lapse microscopy can be used to examine how that protein and the cellular components that it is part of move within the cell during the cell's life cycle. This Methods in Enzymology volume deals with the utility of green fluorescent protein (GFP). The OVID database (including MEDLINE, Current Contents, and other sources) lists nine references to GFP for the ten-year period 1985-1994. In contrast, in less than four years thereafter, over 500 references are listed, a testament to the rapid growth of interest in this probe. This volume documents many diverse uses for this interesting molecule in disciplines that broadly span biology. The methods presented include shortcuts and conveniences not included in previously published sources. The techniques are described in a context that allows comparisons to other related methodologies-such comparisons are valuable to readers who must adapt existing procedures to new systems. Also, so far as possible, methodologies have been presented in a manner that stresses their general applicability and potential limitations. The volume provides a substantial and current overview of the extant methodology in the field and a view of its rapid development. The critically acclaimed laboratory standard for more than forty years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly await

Key Features

@introbul:Key Features @bul:* Monitoring of Physiological Processes * Localization of Molecules * Special Uses * Mutants * Variants of GFP


Biochemists, cell biologists, geneticists, microbiologists, molecular biologists, and biophysicists.

Table of Contents

Monitoring of Physiological Processes: S. Kupzig, S. San Lee and G. Banting, Membrane Trafficking. C. Kaether and H.-H. Gerdes, Monitoring of Protein Secretion with Green Fluorescent Protein. R.M. Hoffman, Green Fluorescent Protein to Visualize Cancer Progression and Metastasis. X. Zhao, T. Duong, C.-C. Huang, S.R. Kain and X. Li, Comparison of Enhanced Green Fluorescent Protein and Its Destabilized Form as Transcription Reporters. S.R. Kain and J.-T. Ma, Early Detection of Apoptosis with Annexin V-Enhanced Green Fluorescent Protein. M. Maniak, Green Fluorescent Protein in the Visualization of Particle Uptake and Fluid-Phase Endocytosis. F. Hanakam and G. Gerisch, Monitoring Intracellular Shuttling of Histidine-rich pH-Sensor Proteins Tagged with Green Fluorescent Protein. S.R. Cronin and R.Y. Hampton, Measuring Protein Degradation with Green Fluorescent Protein. G.L. Hager, Studying Nuclear Receptors with Green Fluorescent Protein Fusions. Localization of Molecules: M.R. Flory and T.N. Davis Localization of Calmodulin in Budding Yeast and Fission Yeast Using Green Fluorescent Protein. K.R. Olson and J.B. Olmsted, Analysis of Microtubule Organization and Dynamics in Living Cells Using Green Fluorescent Protein-MAP4 Chimeras. V. Georget, B. Terouanne, J.-C. Nicholas and C. Sultan, Trafficking of the Androgen Receptor. C.D. Webb and O. Resnekov, Use of Green Fluorescent Protein for Visualization of Cell-Specific Gene Expression and Subcellular Protein Localization in Bacillus subtilis. L.S. Barak, J. Zhang, S.S.G. Ferguson and M.G. Caron, Signaling, Desensitization, and Trafficking of G Protein Coupled Receptor


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Academic Press
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@from:Praise for the Series @qu:"The Methods in Enzymology series represents the gold-standard." @source:--NEUROSCIENCE @qu:"Incomparably useful." @source:--ANALYTICAL BIOCHEMISTRY @qu:"It is a true 'methods' series, including almost every detail from basic theory to sources of equipment and reagents, with timely documentation provided on each page." @source:--BIO/TECHNOLOGY @qu:"The series has been following the growing, changing and creation of new areas of science. It should be on the shelves of all libraries in the world as a whole collection." @source:--CHEMISTRY IN INDUSTRY @qu:"The appearance of another volume in that excellent series, Methods in Enzymology, is always a cause for appreciation for those who wish to successfully carry out a particular technique or prepare an enzyme or metabolic intermediate without the tiresome prospect of searching through unfamiliar literature and perhaps selecting an unproven method which is not easily reproduced." @source:--AMERICAN SOCIETY OF MICROBIOLOGY NEWS @qu:"If we had some way to find the work most often consulted in the laboratory, it could well be the multi-volume series Methods in Enzymology...a great work." @source:--ENZYMOLOGIA @qu:"A series that has established itself as a definitive reference for biochemists." @source:--JOURNAL OF CHROMATOGRAPHY