Dictyostelium discoideum Protocols / Edition 1

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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.

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Product Details

  • ISBN-13: 9781617376771
  • Publisher: Springer-Verlag New York, LLC
  • Publication date: 11/19/2010
  • Series: Methods in Molecular Biology Series, #346
  • Edition description: Softcover reprint of hardcover 1st ed. 2006
  • Edition number: 1
  • Pages: 564
  • Product dimensions: 1.30 (w) x 6.00 (h) x 9.00 (d)

Table of Contents

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 Sk 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

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