Electroporation protocols for microorganisms

Bibliographic Information

Electroporation protocols for microorganisms

edited by Jac A. Nickoloff

(Methods in molecular biology / John M. Walker, series editor, 47)

Humana Press, c1995

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Includes bibliographical references and index

Description and Table of Contents

Description

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.

Table of Contents

Electroporation Theory: Concepts and Mechanisms. Instrumentation. Direct Plasmid Transfer Between Bacterial Species and Electrocuring. Transfer of Episomal and Integrated Plasmids from Saccharomyces cerevisiae to Escherichia coli by Electroporation. Production of cDNA Libraries by Electroporation. Electroporation of RNA into Saccharomyces cerevisiae. Electrofusion of Yeast Protoplasts. Escherichia coli Electrotransformation. Electrotransformation in Salmonella. Electrotransformation of Pseudomonas. Electroporation of Xanthomonas. Transformation of Species with Suicide and Broad Host-Range Plasmids. Electroporation of Francisella tularensis. A Simple and Rapid Method for Transformation of Vibrio Species by Electroporation. Genetic Transformation of Bacteroides spp. Using Electroporation. Electrotransformation of Agrobacterium. Electroporation of Helicobacter pylori. Electrotransformation of Streptococci. Transformation of Lactococcus by Electroporation. Transformation of Lactobacillus by Electroporation. Electrotransformation of Staphylococci. Electroporation and Efficient Transformation of Enterococcus faecalis Grown in High Concentrations of Glycine. Introduction of Recombinant DNA into Clostridium spp. Electroporation of Mycobacteria. Electrotransformation of the Spirochete Borrelia burgdorferi. Yeast Transformation and the Preparation of Frozen Spheroplasts for Electroporation. Ten-Minute Electrotransformation of Saccharomyces cerevisiae. Electroporation of Schizosaccharomyces pombe. Gene Transfer by Electroporation of Filamentous Fungi. Transformation of Candida maltosa by Electroporation. Electroporation of Physarum polycephalum. Electroporation of Dictyostelium discoideum. Gene Transfer by Electroporation of Tetrahymena. Transfection of the African and American Trypanosomes. Electroporation in Giardia lamblia. Index.

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