Green Fluorescent Protein

Since the discovery of the gene for green fluorescent protein (GFP), derived from jellyfish, this protein that emits a green glow has initiated a revolution in molecular biosciences. With this tool, it is now possible to visualize nearly any protein of interest in any cell or tissue of any species. Since the publication of the first edition, there have been tremendously significant technological advances, including development of new mutant variants. Proteins are now available in yellow and blue, and Novel Fluorescent Proteins (NFPs) have expanded their utility in developing biosensors, biological markers, and other biological applications. This updated, expanded new edition places emphasis on the rise of NFPs, including new chapters on NFP properties with detailed protocols, applications of GFPs and NFPs in industry research, and biosensors. This book provides a solid theoretical framework, along with detailed, practical guidance on use of GFPs and NFPs with discussion of potential pitfalls. The expert contributors provide real examples in showing how to tailor GFP/NFP to specific systems, maximize expression, and enhance detection.

This volume is an authoritative and comprehensive treatment of the approaches and techniques used for Green Fluorescent Proteins (GFP). The primary focus of this work is on research using biological systems. The volume covers all aspects of GFP, from its expression in different organisms to specific microscopic and data analysis methods. Key Features * Only volume on Green Fluorescent Protein research * Covers all aspects of GFP * Provides specific microscopic and data analysis methods * Discusses the design and construction of GFP fusion proteins * Covers GFP expression in animals, insects, plants, and microbes * Details procedures for time lapse imaging of living cells * Explains how to implement single molecule fluorescence detection with GFP * Discusses dual label GFP strategies for multicolor fluorescence * Presents fluorescence resonance energy transfer methods with GFPs * Details quantitative fluorescence imaging techniques * Extensively illustrated with color photographs

Over the last ten years, Green Fluorescent Proteins, along with the other spectral variants, have emerged from near obscurity to become a powerful and versatile tool in scientific research. In Viral Applications of Green Fluorescent Protein: Methods and Protocols, leading investigators from around the world contribute detailed examples of both the construction and application of fluorescent proteins delivered by viruses in a format crafted to produce rapid, readily reproducible results. Written in the style of the popular and successful Methods in Molecular BiologyTM series, the chapters include brief introductions to the topics, lists of the necessary materials and reagents, step-by-step laboratory protocols, and Notes sections, which highlight tips on troubleshooting and avoiding known pitfalls. Cutting-edge and easy to use, Viral Applications of Green Fluorescent Protein: Methods and Protocols supplies researchers with an ideal guide to the many uses of GFP and a vital starting point for future studies utilizing this highly adaptable protein.

Since scientists began experimenting with green fluorescent proteins in the middle of the 1990s, these proteins have become one of the most important tools available to researchers in modern medicine and biology. By using them to illuminate other proteins that were previously invisible even under microscope, scientists are now able to observe facets of disease that would have otherwise gone undetected. Green fluorescent proteins are a part of over three million experiments a year, and are invaluable for tasks such as tracking HIV, breeding bird flu-resistant chickens, and confirming the existe ...

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 awaited, frequently consulted, and praised by researchers and reviewers alike. Now with more than 300 volumes (all of them still in print), the series contains much material still relevant today--truly an essential publication for researchers in all fields of life sciences. Key Features * Monitoring of Physiological Processes * Localization of Molecules * Special Uses * Mutants * Variants of GFP

Intended for bench-top use, this lab manual is suitable for both scientists and graduate students, since it combines an update on the most advanced imaging procedures with detailed protocols. Examples, carefully selected from the wide repertoire of cell pyhsiology, cover such different functional aspects as distribution of multiple ions, electrical activity, exo-endocytosis, gene expression, and the cell cycle.

Fluorescent proteins are intimately connected to research in the life sciences. Tagging of gene products with fluorescent proteins has revolutionized all areas of biosciences, ranging from fundamental biochemistry to clinical oncology, to environmental research. The discovery of the Green Fluorescent Protein, its first, seminal application and the ingenious development of a broad palette of fluorescence proteins of other colours, was consequently recognised with the Nobel Prize for Chemistry in 2008. Fluorescent Proteins I is devoted to the basic photophysical and photochemical aspects of fluorescent protein technology. Experienced experts highlight colour tuning, the exploration of switching phenomena and respective methods for their investigation. The book provides a thorough understanding of primary molecular processes allowing the design of fluorescent proteins for specific applications.

Fluorescent proteins are intimately connected to research in the life sciences. Tagging of gene products with fluorescent proteins has revolutionized all areas of biosciences, ranging from fundamental biochemistry to clinical oncology, to environmental research. The discovery of the Green Fluorescent Protein, its first, seminal application and the ingenious development of a broad palette of fluorescence proteins of other colours, was consequently recognised with the Nobel Prize for Chemistry in 2008. Fluorescent Proteins II highlights the physicochemical and biophysical aspects of fluorescent protein technology beyond imaging. It is tailored to meet the needs of physicists, chemists and biologists who are interested in the fundamental properties of fluorescent proteins, while also focussing on specific applications. The implementations described are cutting-edge studies and exemplify how the physical and chemical properties of fluorescent proteins can stimulate novel findings in life sciences.