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.

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.

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. - 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

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.

Methods in Enzymology volumes provide an indispensable tool for the researcher. Each volume is carefully written and edited by experts to contain state-of-the-art reviews and step-by-step protocols. In this volume, we have brought together a number of core protocols concentrating on DNA, complementing the traditional content that is found in past, present and future Methods in Enzymology volumes. - Indispensable tool for the researcher - Carefully written and edited by experts to contain step-by-step protocols - In this volume we have brought together a number of core protocols concentrating on DNA

A comprehensive collection of readily reproducible techniques for the manipulation of recombinant plasmids using the bacterial host E. coli. The authors describe proven methods for cloning DNA into plasmid vectors, transforming plasmids into E. coli, and analyzing recombinant clones. They also include protocols for the construction and screening of libraries, as well as specific techniques for specialized cloning vehicles, such as cosmids, bacterial artificial chromosomes, l vectors, and phagemids. Common downstream applications such as mutagenesis of plasmids, recombinant protein expression, and the use of reporter genes, are also described.

This volume covers a wide range of up-to-date technologies that have been successfully applied to study the chemosensing behavior of the traditional model species, such as Escherichia coli and Salmonella typhimurium, while being also applicable to a wide spectrum of other species. Beginning with an introduction, the sections of the book explore methods for studying bacterial chemotaxis at the population and whole-cell levels, in vivo analysis of receptor function, cryo-EM methods for studying chemoreceptor structure, as well as intracellular movement of chemosensory proteins, high-throughput methods to screen for novel chemoeffectors, and chemical tools and computer simulations for analyzing chemotaxis. Written for the highly successful Methods in Molecular Biology series, chapters include brief introductions to their topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips for troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Bacterial Chemosensing: Methods and Protocols provides an extensive repertoire of approaches that can be extended to understanding chemotaxis, in particular, and chemosensing, in general, in the context of the enormously varied lifestyles adopted in the larger bacterial world.

Methods in Enzymology volumes provide an indispensable tool for the researcher. Each volume is carefully written and edited by experts to contain state-of-the-art reviews and step-by-step protocols. In this volume, we have brought together a number of core protocols concentrating on Cell, Lipid and Carbohydrate, complementing the traditional content that is found in past, present and future Methods in Enzymology volumes. - Indispensable tool for the researcher - Carefully written and edited by experts to contain step-by-step protocols - In this volume we have brought together a number of core protocols concentrating on Cell, Lipid and Carbohydrate