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This detailed volume explores methods currently used to investigate the cell wall of various bacterial species and pathogens. By using a combination of genetic, molecular, biochemical, and cytological techniques, the protocols address many fundamental questions involving the composition, biosynthesis, and regulation of bacterial peptidoglycan. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, as well as tips for troubleshooting and avoiding known pitfalls.
Authoritative and practical, The Bacterial Cell Wall: Methods and Protocols provides current and future researchers with a compilation of many of the most important and useful procedures in a single resource.
List of contents
Bioorthogonal Labeling and Click-Chemistry-Based Visualization of the Tannerella forsythia Cell Wall.- Probing Membrane-Associated Cytoskeletal Oligomers of the Bacterial Divisome by Electron Microscopy and Tomography.- Visualization of a Cell Wall Hydrolase Inhibitor in Fusobacterium nucleatum by Immunofluorescence Microscopy.- Computational and Biophysical Approaches to Identify Cell Wall-Associated Modulators in Salmonella enterica Serovar Typhi.- Employing Cloning-Independent Mutagenesis of Parvimonas micra for the Study of Cell Wall Biogenesis.- A New Method for Gene Deletion to Investigate Cell Wall Biogenesis in Fusobacterium nucleatum.- Super-Resolution Microscopy of the Bacterial Cell Wall Labeled by Fluorescent D-Amino Acids.- Type I Lipoteichoic Acid (LTA) Detection by Western Blot.- Type I Lipoteichoic Acid (LTA) Purification by Hydrophobic Interaction Chromatography and Structural Analysis by 2D Nuclear Magnetic Resonance (NMR) Spectroscopy.- Single-Copy Gene Editing of a Cell Wall-Anchored Pilin in Actinomyces oris.- Quantifying the Kinetics of Pilus-Specific Sortase-Catalyzed Crosslinking Using High-Performance Liquid Chromatography.- Detection of Cell Wall-Anchoring Machinery by Immunogold-Labeling Thin-Section Electron Microscopy.- Localization of the Remnant of a Cell Wall Sorting Signal and Its Interaction with a Sensor Kinase.- Determination of the Crystal Structure of the Cell Wall-Anchored Proteins and Pilins.- Tracking Cell Wall Anchored Proteins in Gram-Positive Bacteria.- Probing Bacterial Cell Division and Cell Envelope Biogenesis with Live-Cell Fluorescence Microscopy.- Assembling the Bacillus subtilis Spore Coat Basement Layer on Spherical Supported Lipid Bilayers.- Structural Determination of Glucosyltransferase C by Cryo-Electron Microscopy.
