Dictyostelium discoideum Protocols
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Dictyostelium discoideum Protocols

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Ludwig Eichinger
845 g
229x152x mm
Vol.346, Methods in Molecular Biology

Part I. The Organism, Community Resources, and Genome-wide StudiesThe Secret Lives of Dictyostelium
Richard H. Kessin
The Genome of Dictyostelium discoideum
Adam Kuspa and William F. Loomis
The cDNA Sequencing Project
Hideko Urushihara, Takahiro Morio, and Yoshimasa Tanaka
dictyBase and the Dicty Stock Center
Petra Fey, Pascale Gaudet, Karen E. Pilcher, Jakob Franke, and Rex L. Chisholm
Analysis of Gene Expression Using cDNA Microarrays
Marcel Kaul and Ludwig Eichinger
Proteomic Analysis of Dictyostelium discoideum
Udo Roth, Stefan Müller, and Franz-Georg Hanisch
Part II. Basic Methods and Molecular Genetic Techniques
Cultivation, Spore Production, and Mating
Hideko Urushihara
Parasexual Genetics Using Axenic Cells
Jason King and Robert Insall
Slug Phototaxis, Thermotaxis, and Spontaneous Turning Behavior
Paul R. Fisher and Sarah J. Annesley
Purification Techniques of Subcellular Compartments for Analytical and Preparative Purposes
Laurence Aubry and Gérard Klein
Generation of Multiple Knockout Mutants Using the Cre-loxP System
Alan R. Kimmel and Jan Faix
Restriction Enzyme-Mediated Integration (REMI) Mutagenesis
Adam Kuspa
RNA Interference and Antisense-Mediated Gene Silencing in Dictyostelium
Markus Kuhlmann, Blagovesta Popova, and Wolfgang Nellen
Part III. Imaging and Localization Methods
Application of Fluorescent Protein Tags as Reporters in Live-Cell Imaging Studies
Annette Müller-Taubenberger
Investigating Gene Expression: In Situ Hybridization and Reporter Genes
Ricardo Escalante and Leandro Sastre
Application of 2D and 3D DIAS to Motion Analysis of Live Cells in Transmission and Confocal Microscopy Imaging
Deborah Wessels, Spencer Kuhl, and David R. Soll
UsingQuantitative Fluorescence Microscopy and FRET Imaging to Measure Spatiotemporal Signaling Events in Single Living Cells
Xuehua Xu, Joseph A. Brzostowski, and Tian Jin
Visualizing Signaling and Cell Movement During the Multicellular Stages of Dictyostelium Development
Dirk Dormann and Cornelis J. Weijer
Under-Agarose Chemotaxis of Dictyostelium discoideum
David Woznica and David A. Knecht
Optimized Fixation and Immunofluorescence Staining Methods for Dictyostelium Cells
Monica Hagedorn, Eva M. Neuhaus, and Thierry Soldati Cryofixation Methods for Ultrastructural Studies of Dictyostelium discoideum
Mark J. Grimson and Richard L. Blanton
Part IV. Dictyostelium as Model Organism
Analysis of Signal Transduction: Formation of cAMP, cGMP, and Ins(1,4,5)P3 In Vivo and In Vitro
Peter J. M. Van Haastert
Assaying Chemotaxis of Dictyostelium Cells
Michelle C. Mendoza and Richard A. Firtel
Characterization of Cross-Linked Actin Filament Gels and Bundles Using Birefringence and Polarized Light Scattering
Ruth Furukawa and Marcus Fechheimer
Quantitative and Microscopic Methods for Studying the Endocytic Pathway
Francisco Rivero and Markus Maniak
Preparation of Intact, Highly Purified Phagosomes from Dictyostelium
Daniel Gotthardt, Régis Dieckmann, Vincent Blancheteau, Claudia Kistler, Frank Reichardt, and Thierry Soldati
Assaying Cell-Cell Adhesion
Salvatore Bozzaro
Periodic Activation of ERK2 and Partial Involvement of G Protein in ERK2 Activation by cAMP in Dictyostelium Cells
Mineko Maeda
An Improved Method for Dictyostelium Centrosome Isolation
Irene Schulz, Yvonne Reinders, Albert Sickmann, and Ralph Gräf
Epigenetics in Dictyostelium
Markus Kaller, Wolfgang Nellen, and Jonathan R. Chubb
Dictyostelium discoideum as a Model to Study
Dictyostelium discoideum is a simple but fascinating eukaryotic microorg- ism, whose natural habitat is deciduous forest soil and decaying leaves, where the amoebae feed on bacteria and grow as independent single cells. Exhaustion of the bacterial food source triggers a developmental program, in which up to 100,000 cells aggregate by chemotaxis towards cAMP. Morphogenesis and cell different- tion then culminate in the production of spores enabling the organism to survive unfavorable conditions. Dictyostelium offers unique advantages for studying f- damental cellular processes with the aid of powerful molecular genetic, bioche- cal, and cell biological tools. These processes include signal transduction, chemotaxis, cell motility, cytokinesis, phagocytosis, and aspects of development such as cell sorting, pattern formation and cell type differentiation. Recently, D- tyostelium was also described as a suitable host for pathogenic bacteria in which one can conveniently study the process of infection. In addition, Dictyostelium has many of the experimental conveniences of Saccharomyces cerevisiae and is pr- ably the best experimentally manipulatable protozoan, providing insight into this diverse group of organisms, which includes some of the most dangerous human parasites. The recent completion of the Dictyostelium genome sequencing project strengthens the position of D. discoideum as a model organism. The completed genome sequence and other valuable community resources constitute the source for basic biological and biomedical research and for genome-wide analyses.