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.

Protocols used in Molecular Biology is a compilation of several examples of molecular biology protocols. Each example is presented with a concise introduction, materials and chemicals required, a step-by-step procedure and troubleshooting tips. Information about the application of the protocol is also provided. The techniques included in this book are essential to research in the fields of proteomics, genomics, cell culture, epigenetic modification and structural biology. The protocols can also be used by clinical researchers (neuroscientists and oncologists, for example) for medical applications (diagnostics, therapeutics and multidisciplinary projects).

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 BiologyTM 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.

Since their discovery in 1869, the dictyostelids have attracted the attention of scientists in a wide variety of fields. This interest has stemmed from their peculiar lifestyle and developmental properties, which were shaped by the evolutionary forces that generated multicellularity during eukaryotic evolution. More recently, the dictyostelids have gained attention due to the striking similarities found at the genomic, cellular and biochemical levels with human cells, which has propelled the species Dictyostelium discoideum to become a model system for biology and medicine in many laboratories. This book covers the latest advances in our knowledge of these extraordinary organisms with topics spanning from their evolutionary history, ecology and diversity to the recent discoveries regarding their cellular and molecular biology.

The volume covers the preparation and analysis of model systems for biological electron microscopy. The volume has chapters about prokaryotic as well as eukaryotic systems that are used as so-called model organisms in modern cell biology. These systems include the most popular systems, such as budding and fission yeast, the roundworm C. elegans, the fly Drosophila, zebrafish, mouse, and Arabidopsis, but also organisms that are less frequently used in cell biology, such as Chlamydomonas, Dictyostelium, Trypanosoma, faltworms, Axolotl and others. In addition, tissues and tissue culture systems are also covered. These systems are used for very diverse areas of cell biology, such as cell division, abscission, intracellular transport, cytoskeletal organization, tissue regeneration and others. Moreover, this issue presents the currently most important methods for the preparation of biological specimens. This volume, however, is not a classic EM methods book. The methods are not the main focus of this issue. The main goal here is to cover the methods in the context of the specific requirements of specimen preparation for each model organism or systems. This will be the first compendium covering the various aspects of sample preparation of very diverse biological systems. - Covers the preparation and analysis of model systems for biological electron microscopy - Includes the most popular systems but also organisms that are less frequently used in cell biology - Presents the currently most important methods for the preparation of biological specimens - First compendium covering the various aspects of sample preparation of very diverse biological systems

Electroporation is one of the most widespread techniques used in modem molecular genetics. It is most commonly used to introduce DNA into cells for investigations of gene structure and function, and in this regard, electroporation is both highly versatile, being effective with nearly all species and cell types, and highly efficient. For many cell types, electroporation is either the most efficient or the only means known to effect gene transfer. However, exposure of cells to brief, hi- intensity electric fields has found broad application in other aspects of biological research, and is now routinely used to introduce other types of biological and analytic molecules into cells, to induce cell-cell fusion, and to transfer DNA directly between different species. The first seven chapters of Electroporation Protocols for Micro organisms describe the underlying theory of electroporation, the com mercially available instrumentation, and a number of specialized electroporation applications, such as cDNA library construction and interspecies DNA electrotransfer. Each of the remaining chapters pre sents a well developed method for electrotransformation of a particular bacterial, fungal, or protist species. These chapters also serve to intro duce those new to the field the important research questions that are currently being addressed with particular organisms, highlighting both the major advantages and limitations of each species as a model organ ism, and explaining the roles that electroporation has played in the development of the molecular genetic systems currently in use.

Dictyostelium Discoideum: Molecular Approaches to Cell Biology