Green Fluorescent Protein

ISBN-13: 9781617372469 / Angielski / Miękka / 2010 / 394 str.

Barry W. Hicks

Opis

Could there be a better time to be a life scientist? In the past two decades, a host of new techniques have been added to the tool chests of biochemists and molecular biologists. A wonderful benefit of the basic scientific research that fueled the advances in these fields is the wide variety of direct applications in agriculture and medicine. Even with all of these advances, and with the accompanying explosion in computer and information technology, it is clear that the depth of our ignorance vastly exceeds the breadth of our knowledge about complex organisms at the molecular level. Any new techniques or materials that allow us to extend our research-based knowledge should be w- comed and utilized to their fullest potentials. With the cloning of the green fluorescent protein (GFP) from Aequorea victoria in 1992, another valuable tool was added to the arsenal. In Green Fluorescent Proteins: Applications and Protocols examples of how GFP can be utilized in a variety of fields are presented. Although the text has chapters that emphasize different areas of s- cialization, it is not meant to send molecular biologists to one section, botanists to another, and clinicians to still another. Perhaps the most valuable exchange for people in any discipline will come from seeing how others have been able to apply GFP in fields outside of their immediate areas of expertise. GFP from Aequorea victoria is a fluorescent marker protein, and there are certainly other useful fluorophore markers.

Recenzje

"The breadth of applications for green fluorescent protein will make this book useful for many laboratories and may provide new approaches for others not previously familiar with gfp as a tool. Gfp has provided a wide ranging and valuable tool for many investigators. This is a very useful set of protocols, even for those who don't think they have an application for the technique. It is especially valuable to have the diverse methods compiled in a single book. Most will find something of value." -Doody's Health Sciences Book Review Journal

"...this book truly is a collection of "Applications and protocols" and such is the degree of detail in the methods described that it should perhaps have been sub-titled "A Laboratory Manual." -The Analyst

Spis treści

Manipulation of Green Fluorescent Protein Structure at the Genetic Level.- Amplification of Representative cDNA Samples from Microscopic Amounts of Invertebrate Tissue to Search for New Genes.- Use of cobA and cysGA as Red Fluorescent Indicators.- Circular Permutation of the Green Fluorescent Protein.- Evolutionary Design of Generic Green Fluorescent Protein Biosensors.- Random Insertion of Green Fluorescent Protein into the Regulatory Subunit of Cyclic Adenosine Monophosphate-Dependent Protein Kinase.- Circular mRNA Encoding for Monomeric and Polymeric Green Fluorescent Protein.- Detection and Imaging of Green Fluorescent Protein.- Fluorescence Lifetime Imaging (FLlM) of Green Fluorescent Fusion Proteins in Living Cells.- 8 Fluorescence Resonance Energy Transfer (FRET) Applications Using Green Fluorescent Protein.- Bioluminescence Resonance Energy Transfer Assays for Protein-Protein Interactions in Living Cells.- Whole-Body Fluorescence Imaging with Green Fluorescence Protein.- Green Fluorescent Protein to Monitor Protein Distribution and Trafficking.- Drug-Induced Translocation of Nucleolar Proteins Fused to Green Fluorescent Protein.- Light-Induced Nuclear Targeting of PhytochromeB-sGreen Fluorescent Protein in Plants.- Mechanisms of Protein Trafficking.- Analysis of Nucleocytoplasmic Transport Using Green Fluorescent Protein.- Green Fluorescent Protein in Transgenic Organisms.- Transgenic Bovine Embryo Selection Using Green Fluorescent Protein.- Development of Glycosylphosphatidylinositol-Anchored Enhanced Green Fluorescent Protein.- Transgenic Zebrafish Expressing Green Fluorescent Protein.- Transgenic Insects Expressing Green Fluorescent Protein-Silk Fibroin Light Chain Fusion Protein in Transgenic Silkworms.- Green Fluorescent Protein inTransgenic Plants.- Green Fluorescent Protein Biosensors.- Green Fluorescent Protein Calcium Biosensors.- Green Fluorescent Protein Fluobody Immunosensors.- 22 Green Fluorescent Protein-Based Protein Kinase Biosensor Substrates.- Green Fluorescent Protein Urea Sensors.- Viral Applications of Green Fluorescent Protein.- Using Green Fluorescent Protein to Monitor Measles Virus Cell-to-Cell Spread by Time-Lapse Confocal Microscopy.- Tracking and Selection of Retrovirally Transduced Murine Bone Marrow Cells Using Green Fluorescent Protein.- Green Fluorescent Protein as a Reporter of Adenovirus-Mediated Gene Transfer and Expression in the Hypothalamic-Neurohypophyseal System.- Enhancement of Green Fluorescent Protein Expression in Adeno-Associated Virus with the Woodchuck Hepatitis Virus Post-Transcriptional Regulatory Element.- Construction of Infectious Simian Varicella Virus Expressing Green Fluorescent Protein.- Green Fluorescent Protein in Retroviral Vector Constructs as Marker and Reporter of Gene Expression for Cell and Gene Therapy Applications.

Blurb

Green fluorescent protein (GFP), a critically important marker protein, allows direct visualization in live samples without the aid of additional sample preparation. In Green Fluorescent Protein: Applications and Protocols, Barry W. Hicks and a team of well-practiced experimentalists demonstrate the power and the versatility of this marker with a cutting-edge collection of readily reproducible GFP techniques. These include methods for detecting and imaging GFP (FRET, BRET, and whole-body imaging), for using GFP to create and select for transgenic organisms, for its use as a biosensor, and for studying viruses. Each method is described in step-by-step detail to ensure robust results and can be used in applications ranging from agriculture and botany to human gene therapy. Extensive notes review the materials required, explain the scientific basis of each step and possible sources of error, and provide tips for overcoming problems.
Highly practical and state-of-the-art, Green Fluorescent Protein: Applications and Protocols makes available to all biological and biomedical experimentalists a full array of the powerful new GFP techniques.