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The Bacterial Flagellum

This volume examines the structure and dynamics of the bacterial flagellum using bacterial genetics, molecular biology, biochemistry, structural biology, biophysics, cell biology, and molecular dynamics simulation. The chapters are divided into 4 parts: Part I describes flagellar type III protein exports, assembly, and gene regulation in S. enterica; Part II explains how to isolate the flagella from the bacterial cell bodies, and further explains how to conduct high-resolution structural and functional analyses of the flagellar motor; Part III talks about how to measure flagellar motor rotation over a wide range of external load, how to measure ion motive force across the cytoplasmic membrane, and how to measure dynamic properties of the flagellar motor proteins by fluorescence microscopy with single molecule precision; and Part IV explores the structure and function of Spirochetal, Vibrio, Shewanella, and Magnetococcus flagellar motors. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and comprehensive, The Bacterial Flagellum: Methods and Protocols aims to provide valuable and vital research to aid in the investigation of the bacterial flagellum resulting from various bacterial species.
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Table of Contents

Preface... Table of Contents... Contributors... Part I Flagellar Type III Protein Export, Assembly and Gene Regulation in Salmonella enterica 1. Fuel of the Bacterial Flagellar Type III Protein Export Apparatus Tohru Minamino, Miki Kinoshita and Keiichi Namba 2. Interactions of Flagellar Structural Subunits with the Membrane Export Components Machinery Lewis D.B. Evans, Paul M. Bergen, Owain J. Bryant and Gillian M. Fraser 3. Fluorescent Microscopy Techniques to Study Hook Length Control and Flagella Formation Marc Erhardt 4. Coupling of Flagellar Gene Expression with Assembly in Salmonella enterica Fabienne F.V. Chevance and Kelly T Hughes 5. Dynamic Measures of Flagellar Gene Expression Santosh Koirala and Christopher V. Rao Part II Structure of the Bacterial Flagellum 6. Purification and Characterization of the Bacterial Flagellar Basal Body from Salmonella enterica Shin-Ichi Aizawa 7. Design and Preparation of the Fragment Proteins of the Flagellar Components Suitable for X-Ray Crystal Structure Analysis Katsumi Imada 8. Structural Analysis of the Flagellar Component Proteins in Solution by Small Angle X-Ray Scattering Lawrence K. Lee 9. Structural Study of the Bacterial Flagellar Basal Body by Electron Cryomicroscopy and Image Analysis Akihoro Kawamoto and Keiichi Namba 10. Structure of the MotA/B Proton Channel Akio Kitao and Yasutaka Nishihara 11. Mechanism of Stator Assembly and Incorporation into the Flagellar Motor Seiji Kojima Part III Dynamics of the Bacterial Flagellar Motor 12. Rotation Measurements of Tethered Cells Yuichi Inoue 13. Tracking the Movement of a Single Prokaryotic Cell in Extreme Environmental Conditions Masayoshi Nishiyama and Yoshiyuki Arai 14. Measurements of the Rotation of the Flagellar Motor by Bead Assay Taishi Kasai and Yoshiyuki Sowa 15. Measurements of Ion-Motive Force Across the Cell Membrane Tsai-Shun Lin, Yi-Ren Sun, and Chien-Jung Lo 16. Stoichiometry and Turnover of the Stator and Rotor Yusuke V. Morimoto and Tohru Minamino 17. Direct Imaging of Intracellular Signaling Molecule Responsible for the Bacterial Chemotaxis Hajime Fukuoka Part IV Structural Diversity of the Bacterial Flagellar Motors Derived from Different Bacterial Species 18. In Situ Structural Analysis of the Spirochetal Flagellar Motor by Cryo-Electron Tomography Shiwei Zhu, Zhuan Qin, Juyu Wang, Dustin R. Morado, and Jun Liu 19. Motility of Spirochetes< Shuichi Nakamura and Md. Shafiqul Islam 20. Structure of the Sodium-Driven Flagellar Motor in Marine Vibrio Yasuhiro Onoue and Michio Homma 21. Chemotactic Behaviors of Vibrio Cholerae Cells Ikuro Kawagishi and So-ichiro Nishiyama 22. Purification of Fla2 Flagella of Rhodobacter sphaeroides Javier de la Mora, Laura Camarena, and Georges Dreyfus 23. Dynamics in the Dual Fuel Flagellar Motor of Shewanella oneidensisâ MR-1 Susanne Brenzinger and Kai M. Thormann 24. Ion Selectivity of the Flagellar Motors Derived from the Alkaliphilic Bacillus and Paenibacillus Species Yuka Takahashi and Masahiro Ito 25. Measurement of Free-Swimming Motility and Magnetotactic Behavior of Magnetococcus massalia Strain MO-1 Wei-Jia Zhang, Sheng-Da Zhang, and Long-Fei Wu

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