Full Description
Dictyostelium discoideum is a well-establish eukaryotic model organism that offers unique advantages for studying fundamental cellular processes, including signal transduction, random and directed cell motility, cytokinesis, endocytosis and vesicle transport and development. Dictyostelium is also increasingly used for the investigation of human disease genes and the crosstalk between host and pathogen. The availability of the genome sequence at a mouse click together with a whole range of supporting information and resources along with a powerful armoury of molecular genetics techniques have considerably enhanced the experimental attractiveness of D. discoideum in recent years.
The second edition of Dictyostelium discoideum Protocols incorporates the most recent developments in a number of fields. The book is divided into four parts. The first part provides an introduction to the amoebozoa and community resources. The second part presents large-scale analysis methods made possible by the completion of the Dictyostelium genome sequence. The third part is dedicated to molecular genetics techniques, cell biological, biochemical and biophysical methods. The chapters in the fourth part describe the use of Dictyostelium as a model system to study vesicle formation, trafficking and infection by bacterial pathogens.
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 key tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Dictyostelium discoideum Protocols, Second Edition seeks to aid scientists in the application of innovating techniques to study a range of fundamental biological processes in this attractive model organism.
Contents
The Amoebozoa.- The Model Organism Dictyostelium discoideum.-Comparative Genomics of the Dictyostelids.- One Stop Shop for Everything Dictyostelium: DictyBase and the Dicty Stock Center in 2012.- Fluorescent Reporters and Methods to Analyze Fluorescent Signals.-Collection and Cultivation of Dictyostelids from the Wild.-Identification and Verification of microRNAs by High-throughput Sequencing.- Transcriptional Profiling of Dictyostelium with RNA Sequencing.- Analysis of Chromatin Organization by Deep Sequencing Technologies.- Pharmacogenetics of Resistance to Cisplatin and Other Anti-cancer Drugs and the Role of Sphingolipid Metabolism.-N-glycomic and -Glycoproteomic Studies in the Social Amoebae.-Measuring Cheating, Fitness, and Segregation in D. discoideum.-The Application of the Cre-loxP System for Generating Multiple Knock-Out and Knock-In Targeted Loci.- Extrachromosomal Inducible Expression.- Isolation of Dictyostelium Nuclei for Light and Electron Microscopy.- Investigation of DNA Repair Pathway Activity.- Transcript Localization in D. discoideum Cells by RNA FISH.- Analysis of Mitochondrial Gene Expression.- Mitochondrial Respiratory Complex Function and the Phenotypic Consequences of Dysfunction.- Micropipette Aspiration for Studying Cellular Mechanosensory Responses and Mechanics .- Quantitative Analysis of Phagocytosis and Phagosome Maturation.- Setting Up and Monitoring an Infection of Dictyostelium discoideum with Mycobacteria.- Isolation of Pathogen-containing Vacuoles.- Immuno-magnetic Purification of Fluorescent Legionella-containing Vacuoles.- Secretory Lysosomes in Dictyostelium: Visualization, Characterization and Dynamics.- Monitoring Autophagy in Dictyostelium.
