Dictyostelium discoideum Protocols

1. Front Matter

   Pages i-xv

   PDF

2. The Organism, Community Resources, and Genome-wide Studies

   1. Front Matter

      Pages 1-1

      PDF

   2. The Secret Lives of Dictyostelium

      • Richard H. Kessin

      Pages 3-13

   3. The Genome of Dictyostelium discoideum

      • Adam Kuspa, William F. Loomis

      Pages 15-29

   4. The cDNA Sequencing Project

      • Hideko Urushihara, Takahiro Morio, Yoshimasa Tanaka

      Pages 31-49

   5. dictyBase and the Dicty Stock Center

      • Petra Fey, Pascale Gaudet, Karen E. Pilcher, Jakob Franke, Rex L. Chisholm

      Pages 51-74

   6. Analysis of Gene Expression Using cDNA Microarrays

      • Marcel Kaul, Ludwig Eichinger

      Pages 75-93

   7. Proteomic Analysis of Dictyostelium discoideum

      • Udo Roth, Stefan Müller, Franz-Georg Hanisch

      Pages 95-109

3. Basic Methods and Molecular Genetic Techniques

   1. Front Matter

      Pages 111-111

      PDF

   2. Cultivation, Spore Production, and Mating

      • Hideko Urushihara

      Pages 113-124

   3. Parasexual Genetics Using Axenic Cells

      • Jason King, Robert Insall

      Pages 125-135

   4. Slug Phototaxis, Thermotaxis, and Spontaneous Turning Behavior

      • Paul R. Fisher, Sarah J. Annesley

      Pages 137-170

   5. Purification Techniques of Subcellular Compartments for Analytical and Preparative Purposes

      • Laurence Aubry, Gérard Klein

      Pages 171-185

   6. Generation of Multiple Knockout Mutants Using the Cre-loxP System

      • Alan R. Kimmel, Jan Faix

      Pages 187-199

   7. Restriction Enzyme-Mediated Integration (REMI) Mutagenesis

      • Adam Kuspa

      Pages 201-209

   8. RNA Interference and Antisense-Mediated Gene Silencing in Dictyostelium

      • Markus Kuhlmann, Blagovesta Popova, Wolfgang Nellen

      Pages 211-226

4. Imaging and Localization Methods

   1. Front Matter

      Pages 227-227

      PDF

   2. Application of Fluorescent Protein Tags as Reporters in Live-Cell Imaging Studies

      • Annette Müller-Taubenberger

      Pages 229-246

   3. Investigating Gene Expression

      • Ricardo Escalante, Leandro Sastre

      Pages 247-260

   4. Application of 2D and 3D DIAS to Motion Analysis of Live Cells in Transmission and Confocal Microscopy Imaging

      • Deborah Wessels, Spencer Kuhl, David R. Soll

      Pages 261-279

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.

• Activation
• DNA sequencing
• Endoplasmatisches Reticulum
• Microarray
• Vivo
• cellular processes
• gene expression
• genes
• hybridization
• signal transduction

• Center for Biochemistry and Center for Molecular Medicine Cologne, University of Cologne, Cologne, Germany

  Ludwig Eichinger, Francisco Rivero

Policies and ethics