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.

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

5. Dynamic Measures of Flagellar Gene Expression
Santosh Koirala and Christopher V. Rao

Part II Structure of the Bacterial Flagellum

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

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

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 oneidensis 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