Dictyostelium discoideum Protocols

Part I. The Organism, Community Resources, and Genome-wide Studies The 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