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.

• First Edition Preface viiPreface xiContributors xiii1. DISCOVERY OF GREEN FLUORESCENT PROTEIN 1Osamu Shimomura2. PHOTONS FOR REPORTING MOLECULAR EVENTS: GREEN FLUORESCENT PROTEIN AND FOUR LUCIFERASE SYSTEMS 15J. Woodland Hastings and James G. Morin3. BIOCHEMICAL AND PHYSICAL PROPERTIES OF GREEN FLUORESCENT PROTEIN 39William W. Ward4. THE THREE-DIMENSIONAL STRUCTURE OF GREEN FLUORESCENT PROTEIN AND ITS IMPLICATIONS FOR FUNCTION AND DESIGN 67George N. Phillips, Jr.5. MOLECULAR BIOLOGY AND MUTATION OF GREEN FLUORESCENT PROTEIN 83David A. Zacharias and Roger Y. Tsien6. DISCOVERY AND PROPERTIES OF GFP-LIKE PROTEINS FROM NONBIOLUMINESCENT ANTHOZOA 121Konstantin A. Lukyanov, Dmitry M. Chudakov, Arkady F. Fradkov, Yulii A. Labas, Mikhail V. Matz, and Sergey Lukyanov7. EVOLUTION OF FUNCTION AND COLOR IN GFP-LIKE PROTEINS 139Mikhail V. Matz, Yulii A. Labas, and Juan Ugalde8. THE USES OF GREEN FLUORESCENT PROTEIN IN PROKARYOTES 163Raphael H. Valdivia, Brendan P. Cormack, and Stanley Falkow9. THE USES OF GREEN FLUORESCENT PROTEIN IN YEASTS 179Amy L. Hitchcock, Jason A. Kahana, and Pamela A. Silver10. USES OF GFP IN CAENORHABDITIS ELEGANS 203Oliver Hobert and Paula Loria11. GREEN FLUORESCENT PROTEIN APPLICATIONS IN DROSOPHILA 227Tulle Hazelrigg and Jennifer H. Mansfield12. THE USES OF GREEN FLUORESCENT PROTEIN IN PLANTS 259Jim Haseloff and Kriby R. Siemering13. USES OF GFP IN TRANSGENIC VERTEBRATES 285Sean Magason, Adam Amsterdam, Nancy Hopkins, and Shuo Lin14. THE USES OF GREEN FLUORESCENT PROTEIN IN MAMMALIAN CELLS 305Theresa H. Ward and Jennifer Lippincott-Schwartz15. PRACTICAL CONSIDERATIONS FOR USE OF REEF CORAL FLUORESCENT PROTEINS IN MAMMALIAN CELLS: APPLICATIONS IN FLUORESCENCE MICROSCOPY AND FLOW CYTOMETRY 339Yu Fang, Olivier Déry, Michael Haugwitz, Pierre Turpin, and Steven R. Kain16. PHARMACEUTICAL APPLICATIONS OF GFP AND RCFP 361Nicola Bevan and Stephen Rees17. REASSEMBLED GFP: DETECTING PROTEIN–PROTEIN INTERACTIONS AND PROTEIN EXPRESSION PATTERNS 391Thomas J. Magliery and Lynne ReganMethods and Protocols 407Steven R. KainIndex 423

"...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.

• First Edition Preface viiPreface xiContributors xiii1. DISCOVERY OF GREEN FLUORESCENT PROTEIN 1Osamu Shimomura2. PHOTONS FOR REPORTING MOLECULAR EVENTS: GREEN FLUORESCENT PROTEIN AND FOUR LUCIFERASE SYSTEMS 15J. Woodland Hastings and James G. Morin3. BIOCHEMICAL AND PHYSICAL PROPERTIES OF GREEN FLUORESCENT PROTEIN 39William W. Ward4. THE THREE-DIMENSIONAL STRUCTURE OF GREEN FLUORESCENT PROTEIN AND ITS IMPLICATIONS FOR FUNCTION AND DESIGN 67George N. Phillips, Jr.5. MOLECULAR BIOLOGY AND MUTATION OF GREEN FLUORESCENT PROTEIN 83David A. Zacharias and Roger Y. Tsien6. DISCOVERY AND PROPERTIES OF GFP-LIKE PROTEINS FROM NONBIOLUMINESCENT ANTHOZOA 121Konstantin A. Lukyanov, Dmitry M. Chudakov, Arkady F. Fradkov, Yulii A. Labas, Mikhail V. Matz, and Sergey Lukyanov7. EVOLUTION OF FUNCTION AND COLOR IN GFP-LIKE PROTEINS 139Mikhail V. Matz, Yulii A. Labas, and Juan Ugalde8. THE USES OF GREEN FLUORESCENT PROTEIN IN PROKARYOTES 163Raphael H. Valdivia, Brendan P. Cormack, and Stanley Falkow9. THE USES OF GREEN FLUORESCENT PROTEIN IN YEASTS 179Amy L. Hitchcock, Jason A. Kahana, and Pamela A. Silver10. USES OF GFP IN CAENORHABDITIS ELEGANS 203Oliver Hobert and Paula Loria11. GREEN FLUORESCENT PROTEIN APPLICATIONS IN DROSOPHILA 227Tulle Hazelrigg and Jennifer H. Mansfield12. THE USES OF GREEN FLUORESCENT PROTEIN IN PLANTS 259Jim Haseloff and Kriby R. Siemering13. USES OF GFP IN TRANSGENIC VERTEBRATES 285Sean Magason, Adam Amsterdam, Nancy Hopkins, and Shuo Lin14. THE USES OF GREEN FLUORESCENT PROTEIN IN MAMMALIAN CELLS 305Theresa H. Ward and Jennifer Lippincott-Schwartz15. PRACTICAL CONSIDERATIONS FOR USE OF REEF CORAL FLUORESCENT PROTEINS IN MAMMALIAN CELLS: APPLICATIONS IN FLUORESCENCE MICROSCOPY AND FLOW CYTOMETRY 339Yu Fang, Olivier Déry, Michael Haugwitz, Pierre Turpin, and Steven R. Kain16. PHARMACEUTICAL APPLICATIONS OF GFP AND RCFP 361Nicola Bevan and Stephen Rees17. REASSEMBLED GFP: DETECTING PROTEIN–PROTEIN INTERACTIONS AND PROTEIN EXPRESSION PATTERNS 391Thomas J. Magliery and Lynne ReganMethods and Protocols 407Steven R. KainIndex 423

"...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.

• First Edition Preface viiPreface xiContributors xiii1. DISCOVERY OF GREEN FLUORESCENT PROTEIN 1Osamu Shimomura2. PHOTONS FOR REPORTING MOLECULAR EVENTS: GREEN FLUORESCENT PROTEIN AND FOUR LUCIFERASE SYSTEMS 15J. Woodland Hastings and James G. Morin3. BIOCHEMICAL AND PHYSICAL PROPERTIES OF GREEN FLUORESCENT PROTEIN 39William W. Ward4. THE THREE-DIMENSIONAL STRUCTURE OF GREEN FLUORESCENT PROTEIN AND ITS IMPLICATIONS FOR FUNCTION AND DESIGN 67George N. Phillips, Jr.5. MOLECULAR BIOLOGY AND MUTATION OF GREEN FLUORESCENT PROTEIN 83David A. Zacharias and Roger Y. Tsien6. DISCOVERY AND PROPERTIES OF GFP-LIKE PROTEINS FROM NONBIOLUMINESCENT ANTHOZOA 121Konstantin A. Lukyanov, Dmitry M. Chudakov, Arkady F. Fradkov, Yulii A. Labas, Mikhail V. Matz, and Sergey Lukyanov7. EVOLUTION OF FUNCTION AND COLOR IN GFP-LIKE PROTEINS 139Mikhail V. Matz, Yulii A. Labas, and Juan Ugalde8. THE USES OF GREEN FLUORESCENT PROTEIN IN PROKARYOTES 163Raphael H. Valdivia, Brendan P. Cormack, and Stanley Falkow9. THE USES OF GREEN FLUORESCENT PROTEIN IN YEASTS 179Amy L. Hitchcock, Jason A. Kahana, and Pamela A. Silver10. USES OF GFP IN CAENORHABDITIS ELEGANS 203Oliver Hobert and Paula Loria11. GREEN FLUORESCENT PROTEIN APPLICATIONS IN DROSOPHILA 227Tulle Hazelrigg and Jennifer H. Mansfield12. THE USES OF GREEN FLUORESCENT PROTEIN IN PLANTS 259Jim Haseloff and Kriby R. Siemering13. USES OF GFP IN TRANSGENIC VERTEBRATES 285Sean Magason, Adam Amsterdam, Nancy Hopkins, and Shuo Lin14. THE USES OF GREEN FLUORESCENT PROTEIN IN MAMMALIAN CELLS 305Theresa H. Ward and Jennifer Lippincott-Schwartz15. PRACTICAL CONSIDERATIONS FOR USE OF REEF CORAL FLUORESCENT PROTEINS IN MAMMALIAN CELLS: APPLICATIONS IN FLUORESCENCE MICROSCOPY AND FLOW CYTOMETRY 339Yu Fang, Olivier Déry, Michael Haugwitz, Pierre Turpin, and Steven R. Kain16. PHARMACEUTICAL APPLICATIONS OF GFP AND RCFP 361Nicola Bevan and Stephen Rees17. REASSEMBLED GFP: DETECTING PROTEIN–PROTEIN INTERACTIONS AND PROTEIN EXPRESSION PATTERNS 391Thomas J. Magliery and Lynne ReganMethods and Protocols 407Steven R. KainIndex 423

"...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)