细胞技术

The modification of cell surface proteins by plasma membrane and soluble proteases is important for physiological and pathological processes. Methods to identify shed and soluble substrates are crucial to further define the substrate repertoire, termed the substrate degradom ...

The discovery of novel target antigens for antibody-based immunotherapy is still a major challenge. Antibody phage display is one of the technologies that is widely applied for the identification of novel cell surface molecules on intact eukaryotic cells and many reports describe the is ...

We identify and describe a set of tools readily available for integral membrane protein prediction. These tools address two problems: finding potential transmembrane proteins in a pool of new sequences, and identifying their transmembrane regions. All methods involve comparing the ...

The social amoeba Dictyostelium discoideum is already known as a model organism for a variety of cellular and molecular studies. Now that the genome sequencing project has been completed and different tools with which to overexpress or knock out genes are available, this species has also moved ...

A DNA microarray consists of an orderly arrangement of DNA, cDNA, or oligonucleotide probes that represent individual genes of an organism. Today, microarrays are the most important tools used to analyze gene transcription on a large scale. Investigations reach from finding key genes in wh ...

In this chapter, we describe how to conveniently demonstrate, assess, and study cell death in Dictyostelium through simple cell culture, clonogenic tests, and photonic (with the help of staining techniques) and electronic microscopy. Cell death can be conveniently generated using mi ...

dictyBase is the model organism database that houses all the sequence and associated data for Dictyostelium discoideum, including literature, researchers, and strains from the Dicty Stock Center. The database makes it possible to connect genes, proteins, and publications and is desi ...

Dictyostelium has a good potential to serve as a model for the study of chromatin function and epigenetic gene regulation. The organism bridges the complexity of higher eukaryotic systems and the simplicity of yeast in that it harbors pathways that are similar to the former and is accessible to ge ...

The Dictyostelium dicoideum centrosome consists of a box-shaped, layered core structure surrounded by dense nodules embedded in amorphous material, which make up the so-called corona. Thus, it differs markedly from centriole-containing centrosomes in animal cells or the plaque s ...

A major feature in Dictyostelium development is the transition from the unicellular to the multicellular stage, a process brought about by chemotaxis and cell-cell adhesion. Growth-phase cells are weakly cohesive, whereas aggregation-competent cells adhere strongly to each oth ...

Phagocytosis plays a fundamental role in the immune system for the defense against invading microorganisms and the clearing of apoptotic and cancerous cells. The common amoeba Dictyostelium discoideum is a recognized model for professional immune phagocytes and is now commonly us ...

Endocytosis is a process that is essential to the life of all eukaryotic cells. Laboratory strains of Dictyostelium are extremely efficient in the uptake of both particles and fluid. Many different cellular processes feed into the endocytic pathway, and many organelle-associated and cy ...

Fundamental processes in the life of Dictyostelium, such as locomotion, endocytosis, cytokinesis, and morphogenesis, are mediated by the actin cytoskeleton, which is composed of actin, myosins, and numerous actin-binding proteins. An understanding of these processes at the molec ...

Both prokaryote and eukaryote cells can sense and move up chemical concentration gradients (chemotax). As a means of finding food sources during vegetative growth, Dictyostelium discoideum naturally chemotaxes toward chemicals released by bacteria. As part of its developmental ...

Cryopreservation methods, including rapid freezing, freeze-substitution, and low-temperature embedment, lead to superior ultrastructural preservation compared with traditional fixation procedures. This is particularly true for the multicellular stages of Dict ...

Recent years have seen a powerful revival of fluorescence microscopy techniques, both to observe live cells and fixed objects. The limits of sensitivity, simultaneous detection of multiple chromophores, and spatial resolution have all been pushed to the extreme. Therefore, it is essen ...

In the vegetative state, Dictyostelium amoebae are chemotactic toward pterins released by bacteria, whereas during multicellular development, they become chemotactic to endogenously produced cAMP. A variety of assays have been used to visualize and quantify chemotactic move ...

The Dictyostelium discoideum cDNA sequencing project started in 1995, preceding the genome sequencing project. Altogether, 14 cDNA libraries, including full-length ones, were constructed from five different stages of growth and asexual and sexual development, from which near ...

Time-lapse microscopy provides a powerful tool with which to study cell behavior during Dictyostelium development. On a macroscopic level, the overall cell movement patterns that give rise to the complex multicellular structures such as slugs and fruiting bodies can be studied toget ...

The mechanisms that mediate how migratory eukaryotic cells amplify a shallow, extracellular chemoattractant gradient into a steep intracellular gradient of signaling components to guide chemotaxis remains unknown. To unravel these mechanisms, it is essential to quantitat ...