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.Eigenschaften und Anwendungen von nicht fluoreszierenden und von grun fluoreszierenden Proteinen (NFPs bzw. GFPs) stehen im Mittelpunkt dieses Bandes, der auf solider theoretischer Grundlage Einsatzgebiete in der Industrie (etwa in der Biosensorentechnik) diskutiert. Die Beitrage, von ausgewiesenen Experten verfasst, beschreiben das ma?geschneiderte Design von GFPs und NFPs fur spezifische Systeme, die Maximierung der Expression, Ma?nahmen zur Verbesserung der Detektion und andere interessante Themen. Neben vielen praktischen Hinweisen erhalten Sie auch Tipps zum Vermeiden haufiger Fehler. First Edition Preface.

Preface.

Contributors.

1. Discovery of Green Fluorescent Protein (Osamu Shimomura).

2. Photons for Reporting Molecular Events: Green Fluorescent Protein and Four Luciferase Systems (J. Woodland Hastings and James G. Morin).

3. Biochemical and Physical Properties opf Green Fluorescent Protein (William W. Ward).

4. The Three-Dimensional Structure of Green Fluorescent Protein and Its Implications for Function and Design (George N. Phillips).

5. Molecular Biology and Mutation of Green Fluorescent Protein (David A. Zacharias and Roger Y. Tsien).

6. Discovery and Properties of GFP-Like Proteins from Nonbioluminescent Anthozoa (Konstantin A. Lukyanov, Dmitry M. Chudakov, Arkady F. Fradkov, Yulii A. Labas, Mikhail V. Matz, and Sergey Lukyanov).

7. Evolution of Function and Color in GPF-Like Proteins (Mikhail V. Matz, Yulii A. Labas, and Juan Ugalde).

8. The Uses of Green Fluorescent Protein in Prokaryotes (Raphael H. Valdivia, Brendan P. Cormack, and Stanley Falkow).

9. The uses of Green Fluorescent Protein in Yeasts (Amy L. Hitchcock, Jason A. Kahana, and Pamela A. Silver).

10. Uses of GFP in Caenorhabditis Elegans (Oliver Hobert and Paula Loria).

11. Green Fluorescent Protein Applications in Drosophila (Tulle Hazelrigg and Jennifer H. Mansfield).

12.The Uses of Green Fluorescent Protein in Plants (Jim Haseloff and Kirby R. Siemering).

13. Uses of GFP in Transgenic Vertebrates (Sean Magason, Adam Amsterdam, Nancy Hopkins, and Shuo Lin).

14. The Uses of Green Fluorescent Protein in Mammalian Cells (Theresa H. Ward and Jennifer Lippincott-Schwartz).

15. Practical Considerations for Use of Reef Coral Fluorescent Proteins in Mammalian Cells: Applications in Fluorescent Microscopy and Flow Cytometry (Yu Fang, Olivier Déry, Michael Haugwitz, Pierre Turpin, and Steven R. Kain).

16. Pharmaceutical Applications of GFP and RCFP (Nicola Bevan and Stephen Rees).

17. Reassembled GFP: Detecting Protein-Protein Interactions and Protein Expression Patterns (Thomas J. Magliery and Lynne Regan).

Methods and Protocols (Steven R. Kain).

Index. "...a magnificent text that will immensely benefit students and scientists from many biomedical areas." (Annals of Biomedical Engineering, August 2006)

"Any researcher, graduate student or educator with an interest in using GFP should read this book. It will become the 'bible for GFP' users." (The Quarterly Review of Biology, June 2006)

MARTIN CHALFIE, PhD, is William R. Kenan Jr. Professor of Biological Sciences at Columbia University. He is world renowned for introducing the use of GFP as a biological marker. A member of the National Academy of Sciences, Dr. Chalfie serves on the editorial boards of Genome Biology and Molecular Biology of the Cell.

STEVEN R. KAIN, PhD, manages the Applications Gene Expression Group for Agilent Technologies. While director of cell biology at Clontech Laboratories, he was responsible for commercializing the first products based on GFP and produced the popular EGFP variant. He has twelve years' experience with a variety of biotechnology firms in the San Francisco Bay Area.

The latest edition of the definitive text in the field of fluorescent proteins

Green fluorescent protein (GFP), a biological marker cloned from jellyfish, has emerged from virtual obscurity to the forefront of biomolecular research in just a few years. GFP and related fluorescent proteins (FPs) have been intensely studied—the subjects of hundreds of in vivo projects worldwide. These studies have resulted in new applications and published articles, emerging at a breathtaking rate. The success of FPs and their many uses promise to make them some of the most powerful tools in biotechnology, pharmaceutical science, and molecular and cellular biology.

Written by pioneers in the field, this text tackles both theory and practice, offering numerous case studies, examples, illustrations, and troubleshooting tips. It examines how FPs are tailored for specific systems and used to maximize expression, and how variants are generated with altered properties. The text also explores new ways to use FPs and methods for enhancing detection in a variety of organisms and cell types.

This updated and expanded edition places emphasis on the rise of FPs and their applications in industry and biosensor research. Green Fluorescent Protein: Properties, Applications, and Protocols, Second Edition provides: New chapters on FPs, biosensors, and advances in the use of FPs in the biotechnology and pharmaceutical industries Detailed information about protocols using FPs Examples from recent notable studies Background on the biochemical and physical properties, three-dimensional structure, and molecular biology and mutation of FPs The application of FPs across organisms

Green Fluorescent Protein: Properties, Applications, and Protocols, Second Edition is essential reading for researchers and graduate students in molecular and cellular biology, physiology, biochemistry, developmental biology, neuroscience, ecology, in vivo biology, and plant molecular biology.