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In recent years, molecular microbiology has emerged as a top, cutting-edge biological discipline, thanks to the multi-disciplinary and integrative approaches taken by investigators seeking to understand the intricacies of the microbial world and how it affects human health and the biosphere. In Bacterial Cell Surfaces: Methods and Protocols, recent advances in structural biology, proteomics, and imaging techniques, together with the traditional biochemical and genetic approaches, are provided in order to present an exciting look into the structure, function, and regulation of the bacterial cell envelope. The detailed volume contains examples of traditional and innovative tools for the study of protein structure and function and enzymatic activities, the purification and analysis of macromolecules and their complexes, and the investigation of regulatory mechanisms and cell biological processes. Written in the highly successful Methods in Molecular Biology(TM) 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.
Authoritative and practical, Bacterial Cell Surfaces: Methods and Protocols is aimed at the microbiologist, biochemist, or cell biologist, whether a beginning graduate student or a veteran experimentalist, who wishes to learn new methodologies and take advantage of the years of research and protocol optimization from the best laboratories around the world.
Sommario
A Bird's Eye View of the Bacterial Landscape.- Visualizing the Bacterial Cell Surface: An Overview.- Purification of the Outer Membrane Usher Protein and Periplasmic Chaperone-Subunit Complexes from the P and Type 1 Pilus Systems.- Experimental Manipulation of the Microbial Functional Amyloid Called Curli.- Visualization of Gram-Positive Bacterial Pili.- Single Cell Microfluidic Studies of Bacterial Motility.- Chromatographic Analysis of the Escherichia coli Polysialic Acid Capsule.- Analysis of Exopolysaccharides in Myxococcus xanthus Using Confocal Laser Scanning Microscopy.- Assessment of Multidrug Efflux Assemblies by Surface Plasmon Resonance.- Fluorescence Microscopy and Proteomics to Investigate Subcellular Localization, Assembly, and Function of the Type II Secretion System.- Pore Formation by T3SS Translocators: Liposome Leakage Assay.- Isolation of Bacterial Type IV Machine Subassemblies.- Production and Crystallization of Bacterial Type V Secretion Proteins.- Assembly of Bacterial Outer Membrane Proteins.- The Outer Membrane of Gram-Negative Bacteria: Lipid A Isolation and Characterization.- Quantitative and Qualitative Preparations of Bacterial Outer Membrane Vesicles.- In Vitro Peptidoglycan Synthesis Assay with Lipid II Substrate.- Extraction of Cell Wall-Bound Teichoic Acids and Surface Proteins from Listeria monocytogenes.- Subfractionation and Analysis of the Cell Envelope (Lipo)polysaccharides of Mycobacterium tuberculosis.- Protein Disulfide Bond Formation in the Periplasm: Determination of the In Vivo Redox State of Cysteine Residues.- Using Reporter Genes and the Escherichia coli ASKA Over-Expression Library in Screens for Regulators of the Gram Negative Envelope Stress Response.- Isolation of Bacteria Envelope Proteins.- Patch Clamp Electrophysiology for the Study of Bacterial Ion Channels inGiant Spheroplasts of E. coli.- Electrophysiological Characterization of Bacterial Pore-Forming Proteins in Planar Lipid Bilayers.
Riassunto
In recent years, molecular microbiology has emerged as a top, cutting-edge biological discipline, thanks to the multi-disciplinary and integrative approaches taken by investigators seeking to understand the intricacies of the microbial world and how it affects human health and the biosphere. In Bacterial Cell Surfaces: Methods and Protocols, recent advances in structural biology, proteomics, and imaging techniques, together with the traditional biochemical and genetic approaches, are provided in order to present an exciting look into the structure, function, and regulation of the bacterial cell envelope. The detailed volume contains examples of traditional and innovative tools for the study of protein structure and function and enzymatic activities, the purification and analysis of macromolecules and their complexes, and the investigation of regulatory mechanisms and cell biological processes. 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.
Authoritative and practical, Bacterial Cell Surfaces: Methods and Protocols is aimed at the microbiologist, biochemist, or cell biologist, whether a beginning graduate student or a veteran experimentalist, who wishes to learn new methodologies and take advantage of the years of research and protocol optimization from the best laboratories around the world.
