Translation Initiation: Extract Systems And Molecular Genetics

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For over fifty years the Methods in Enzymology series has been the critically aclaimed laboratory standard and one of the most respected publications in the field of biochemistry. The highly relevant material makes it an essential publication for researchers in all fields of life and related sciences. This volume, the first of three on the topic of Translation Initiation includes articles written by leaders in the field.

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METHODS IN ENZYMOLOGY Editors-in-Chief JOHN N. ABELSON AND MELVIN I. SIMON Division of Biology California Institute of Technology Pasadena, California Founding Editors SIDNEY P. COLOWICK AND NATHAN O. KAPLAN CONTENTS Contributors Preface Volumes in Series 1. Transient Kinetics, Fluorescence, and FRET in Studies of Initiation of Translation in Bacteria xiii xix xxi 1 Pohl Milon, Andrey L. Konevega, Frank Peske, Attilio Fabbretti, Claudio O. Gualerzi, and Marina V. Rodnina 1. Introduction 2. Experimental Outline 3. Materials 4. Experimental Procedures 5. Rapid Kinetic Measurements 6. Quench-Flow Measurements 7. GTPase Activity 8. Dipeptide Formation 9. Applications of the Method 10. Binding of fMet-tRNA to the 30S Subunit 11. FRET to IF3 12. Nucleotide Binding to IF2 13. Subunit Joining Acknowledgments References 2. Binding of mRNA to the Bacterial Translation Initiation Complex 2 5 9 10 20 21 22 22 23 23 23 25 25 26 26 31 Sean M. Studer and Simpson Joseph 1. Introduction 2. Experimental Procedures Acknowledgments References 32 33 43 43 v vi Contents 3. Real-Time Dynamics of Ribosome-Ligand Interaction by Time-Resolved Chemical Probing Methods 45 Attilio Fabbretti, Pohl Milon, Anna Maria Giuliodori, Claudio O. Gualerzi, and Cynthia L. Pon 1. Introduction 2. General Strategy
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