Electrotransformation Of Bacteria

In this manual, protocols for the transformation of about 40 strains of bacteria are described, with the emphasis placed on the individual critical procedural steps, since the practical details mainly depend on the bacterial strain under investigation. This presentation together with the theoretical introductionary chapters, allows users to modify and adapt each protocol to their own experiments. Bacterial strains with relevance in the food industry, biotechnology, medical and veterinary fields, agroindustry and environmental sciences are covered.

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.

This major reference work is a one-shot knowledge base on electroporation and the use of pulsed electric fields of high intensity and their use in biology, medicine, biotechnology, and food and environmental technologies. The Handbook offers a widespread and well-structured compilation of 156 chapters ranging from the foundations to applications in industry and hospital. It is edited and written by most prominent researchers in the field. With regular updates and growing in its volume it is suitable for academic readers and researchers regardless of their disciplinary expertise, and will also be accessible to students and serious general readers. The Handbook's 276 authors have established scholarly credentials and come from a wide range of disciplines. This is crucially important in a highly interdisciplinary field of electroporation and the use of pulsed electric fields of high intensity and its applications in different fields from medicine, biology, food processing, agriculture, process engineering, energy and environment. An Editorial Board of distinguished scholars from across the world has selected and reviewed the various chapters to ensure the highest quality of this Handbook. The book was edited by an international team of Section Editors: P. Thomas Vernier, Boris Rubinsky, Juergen Kolb, Damijan Miklavcic, Marie-Pierre Rols, Javier Raso, Richard Heller, Gregor Serša, Dietrich Knorr, and Eugene Vorobiev.

Cells can be funny. Try to grow them with a slightly wrong recipe, and they turn over and die. But hit them with an electric field strong enough to knock over a horse, and they do enough things to justify international meetings, to fill a sizable book, and to lead one to speak of an entirely new technology for cell manipulation. The very improbability of these events not only raises questions about why things happen but also leads to a long list of practical systems in which the application of strong electric fields might enable the merger of cell contents or the introduction of alien but vital material. Inevitably, the basic questions and the practical applications will not keep in step. The questions are intrinsically tough. It is hard enough to analyze the action of the relatively weak fields that rotate or align cells, but it is nearly impossible to predict responses to the cell-shredding bursts of electricity that cause them to fuse or to open up to very large molecular assemblies. Even so, theoretical studies and systematic examination of model systems have produced some creditable results, ideas which should ultimately provide hints of what to try next.

In Pseudomonas aeruginosa, expert researchers in the field detail many of the methods which are now commonly used to study this fascinating microorganism. Chapters include microbiological methods to high-throughput molecular techniques that have been developed over the last decade. 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, Pseudomonas aeruginosa aids in the continuing study of new and cutting edge findings.

Electroporation is an efficient method to introduce macromolecules such as DNA into a wide variety of cells. Electrofusion results in the fusion of cells and can be used to produce genetic hybrids or hybridoma cells. Guide to Electroporation and Electrofusion is designed to serve the needs of students, experienced researchers, and newcomers to the field. It is a comprehensive manual that presents, in one source, up-to-date, easy-to-follow protocols necessary for efficient electroporation and electrofusion of bacteria, yeast, and plant and animal cells, as well as background information to help users optimize their results through comprehension of the principles behind these techniques. Key Features * Covers fundamentals of electroporation and electrofusion in detail * Molecular events * Mechanisms * Kinetics * Gives extensive practical information * The latest applications * Controlling parameters to maximize efficiency * Available instrumentation * Presents applications of electroporation and electrofusion in current research situations * State-of-the-art modifications to electrical pulses and generators * Application of electroporation and electrofusion to unique, alternative cell and tissue types * Gives straightforward, detailed, easy-to-follow protocols for * Formation of human hybridomas * Introduction of genetic material into plant cells and pollen * Transfection of mammalian cells * Transformation of bacteria, plants, and yeast * Production of altered embryos * Optimization of electroporation by using reporter genes * Comprehensive and up-to-date * Convenient bench-top format * Approximately 125 illustrations complement the text * Complete references with article titles * Written by leading authorities in electroporation and electrofusion

The field of bacterial genetics has been restricted for many years to Escherichia coli and a few other genera of aerobic or facultatively anaerobic bacteria such as Pseudomonas, Bacillus, and Salmonella. The prevailing view up to recent times has been that anaerobic bacteria are interesting organisms but nothing is known about their genetics. To most microbiologists, anaerobic bacteria appeared as a sort of distant domain, reserved for occasional intrusions by taxonomists and medical microbiologists. By the mid-1970s, knowledge of the genetics and molecular biology of anaerobes began to emerge, and then developed rapidly. but also im This was the result of advances in molecular biology techniques, portantly because of improvements in basic techniques for culturing anaerobes and for understanding their biochemistry and other areas of in terest. Investigations in this field were also stimulated by a renewal of interest in their ecology, their role in pathology and in biotransformations, and in the search for alternative renewable sources of energy. The initial idea for this book came from Thomas D. Brock. When Dr. Brock requested my opinion about two years ago on the feasibility of publishing a book on the genetics of anaerobic bacteria, as a part of the Brock/Springer Series in Contemporary Bioscience, I answered positively but I was apprehen sive about assuming the role of editor. However, I was soon reassured by the enthusiastic commitment of those I approached to contribute. Eventually, thanks to the caring cooperation of the contributors, the task became relatively easy.