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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.
List of contents
Fuel of the Bacterial Flagellar Type III Protein Export Apparatus.- Interactions of Flagellar Structural Subunits with the Membrane Export Components Machinery.- Fluorescent Microscopy Techniques to Study Hook Length Control and Flagella Formation.- Coupling of Flagellar Gene Expression with Assembly in Salmonella enterica.- Dynamic Measures of Flagellar Gene Expression.- Purification and Characterization of the Bacterial Flagellar Basal Body from Salmonella enterica.- Design and Preparation of the Fragment Proteins of the Flagellar Components Suitable for X-Ray Crystal Structure Analysis.- Structural Analysis of the Flagellar Component Proteins in Solution by Small Angle X-Ray Scattering.- Structural Study of the Bacterial Flagellar Basal Body by Electron Cryomicroscopy and Image Analysis.- Structure of the MotA/B Proton Channel.- Mechanism of Stator Assembly and Incorporation into the Flagellar Motor.- RotationMeasurements of Tethered Cells.- Tracking the Movement of a Single Prokaryotic Cell in Extreme Environmental Conditions.- Measurements of the Rotation of the Flagellar Motor by Bead Assay.- Measurements of Ion-Motive Force Across the Cell Membrane.- Stoichiometry and Turnover of the Stator and Rotor.- Direct Imaging of Intracellular Signaling Molecule Responsible for the Bacterial Chemotaxis.- In Situ Structural Analysis of the Spirochetal Flagellar Motor by Cryo-Electron Tomography.- Motility of Spirochetes.- Structure of the Sodium-Driven Flagellar Motor in Marine Vibrio.- Chemotactic Behaviors of Vibrio Cholerae Cells.- Purification of Fla2 Flagella of Rhodobacter sphaeroides.- Dynamics in the Dual Fuel Flagellar Motor of Shewanella oneidensis MR-1.- Ion Selectivity of the Flagellar Motors Derived from the Alkaliphilic Bacillus and Paenibacillus Species.- Measurement ofFree-Swimming Motility and Magnetotactic Behavior of Magnetococcus massalia Strain MO-1.
Summary
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.
