实验基础

DNA is constantly being damaged from endogenous and exogenous sources and efficient repair of different types of DNA lesions is essential for the survival of the organism. Dictyostelium is highly resistant to DNA damage and its genome sequence has revealed the presence of multiple repair p ...

The significance of amoebae for studies of nuclear architecture has considerably increased in the recent years. The availability of a protocol for isolation of nuclei in a quality sufficient for high-resolution light and electron microscopy is a prerequisite for such studies. Here we pr ...

Inducible expression systems are very convenient for proteins that induce strong side effects such as retardation of growth or development and are essential for the expression of toxic proteins. In this chapter we describe the doxycycline-inducible expression system, optimized f ...

Dictyostelium provides a well established model system for the study of mitochondrial biology and disease. Mitochondrial dysfunction in Dictyostelium has been generated by knockout of nonessential nuclear genes encoding mitochondrial proteins, by knockout of targeted mit ...

Dictyostelium provides a well-established model system for the study of mitochondrial biology and disease. A complete mitochondrial transcription and RNA-processing map has been generated, while the start site for transcription and the responsible RNA polymerase have also been ...

Biogenesis of ribosomal RNA (rRNA) takes place preliminary in the nucleolus of eukaryotic cells, the site of rDNA transcription. Several processing steps of rRNA molecules have been implied to take place in the cytoplasm. To follow these processing events we have adapted protocols for fluo ...

The model organism Dictyostelium discoideum is a member of the Amoebozoa, one of the six major �divisions of eukaryotes. Amoebozoa comprise a wide variety of amoeboid and flagellate organisms with single cells measuring from 5 μm to several meters across. They have adopted many different li ...

Phagocytosis and phagosome maturation lead to killing and digestion of bacteria by protozoans and innate immune phagocytes. Phagocytosis of particles expressing or coupled to various fluorescent reporters and sensors can be used to monitor quantitatively various parameters ...

Micropipette aspiration (MPA) is a widely applied method for studying cortical tension and deformability. Based on simple hydrostatic principles, this assay allows the application of a specific magnitude of mechanical stress on cells. This powerful method has revealed insights ab ...

Mycobacterium marinum is the causative agent of fish and amphibian tuberculosis in the wild. It is a genetically close cousin of Mycobacterium tuberculosis, and thereby the infection process remarkably shares many of the hallmarks of M. tuberculosis infection in human, at both the cellul ...

Dictyostelium discoideum cells are “professional phagocytes,” as they ingest a large variety of bacteria, yeast, and inert particles. Several bacterial pathogens are able to survive intracellularly within specialized vacuoles of D. discoideum by interfering with host signali ...

Autophagy is an intracellular degradation mechanism essential for cell survival and maintenance of cellular homeostasis, differentiation, and development. Recent research has highlighted the impact of autophagy in neurodegenerative diseases and aging. We are still far from ...

Dictyostelium discoideum is a unicellular eukaryotic model largely used for the study of the endocytic pathway, mainly due to its high resemblance to the mammalian pathway. Over the last years, Dictyostelium has been increasingly employed for investigating the biogenesis and secre ...

Protozoa are natural reservoirs of the environmental bacterium Legionella pneumophila. Upon inhalation of Legionella-laden aerosols, the amoeba-resistant bacteria replicate within human alveolar macrophages causing the severe pneumonia “Legionnaires’ disease.” ...

Much of our knowledge of molecular cellular functions is based on studies with a few number of model organisms that were established during the last 50 years. The social amoeba Dictyostelium discoideum is one such model, and has been particularly useful for the study of cell motility, chemotaxis, ...

The complete genomes of Dictyostelium discoideum, Dictyostelium purpureum, Polysphondylium pallidum and Dictyostelium fasciculatum have been sequenced. The proteins predicted to be encoded by the genes in each species have been compared to each other as well as to the complete com ...

dictyBase (http://dictybase.org), the model organism database for Dictyostelium discoideum, includes the complete genome sequence and expression data for this organism. Relevant literature is integrated into the database, and gene models and functional annotation are manu ...

Dictyostelium discoideum is a commonly used model organism for the study of biological processes such as chemotaxis, cell communication, and development. While these studies primarily focus on a single clone, recent work has revealed a host of questions that can only be answered from studi ...

The use of fluorescent reporters and the development of new imaging technologies have revolutionized studies in cell biology. During recent years the number of fluorescent proteins offering the ability to visualize the distribution of proteins, organelles, and cells has increased ...

High-throughput sequencing methods have become invaluable for detection and analysis of small RNAs. The results are millions of sequences that need to be carefully analyzed by computational methods and preferentially verified by different experimental techniques. Here we des ...