Genetic Recombination: Reviews and Protocols

ISBN-13: 9781588292360 / Angielski / Twarda / 2004 / 260 str.

Alan S. Waldman

Opis

Genetic recombination, in the broadest sense, can be defined as any process in which DNA sequences interact and undergo a transfer of information, producing new recombinant sequences that contain information from each of the original molecules. All organisms have the ability to carry out recombination, and this striking universality speaks to the essential role recombination plays in a variety of biological processes fundamentally important to the maintenance of life. Such processes include DNA repair, regulation of gene expression, disease etiology, meiotic chromosome segregation, and evolution. One important aspect of recombination is that it typically occurs only between sequences that display a high degree of sequence identity. The stringent requirement for homology helps to ensure that, under normal circumstances, a cell is protected from deleterious rearrangements since a swap of genetic information between two nearly identical sequences is not expected to dramatically alter a genome. Recombination between dissimilar sequences, which does happen on occasion, may have such harmful consequences as chromosomal translocations, deletions, or inversions. For many organisms, it is also important that recombination rates are not too high lest the genome become destabilized. Curiously, certain organisms, such as the trypanosome parasite, actually use a high rate of recombination at a particular locus in order to switch antigen expression continually and evade the host immune system effectively."

Spis treści

Part I. Studying Recombination Events in Eukaryotes Determination of Mitotic Recombination Rates by Fluctuation Analysis in Saccharomyces cerevisiae Rachelle Miller Spell and Sue Jinks-Robertson Determination of Intrachromosomal Recombination Rates in Cultured Mammalian Cells Jason A. Smith and Alan S. Waldman Intrachromosomal Homologous Recombination in Arabidopsis thaliana Waltraud Schmidt-Puchta, Nadiya Orel, Anzhela Kyryk, and Holger Puchta Analysis of Recombinational Repair of DNA Double-Strand Breaks in Mammalian Cells With I-SceI Nuclease Jac A. Nickoloff and Mark A. Brenneman Transformation of Monomorphic and Pleomorphic Trypanosoma brucei Richard McCulloch, Erik Vassella, Peter Burton, Michael Boshart, and J. David Barry Forward Genetic Screens for Meiotic and Mitotic Recombination-Defective Mutants in Mice Laura Reinholdt, Terry Ashley, John Schimenti, and Naoko Shima Part II. Recombination as a Reporter of Genomic Instability Detecting Carcinogens With the Yeast DEL Assay Richard J. Brennan and Robert H. Schiestl In Vivo DNA Deletion Assay to Detect Environmental and Genetic Predisposition to Cancer Ramune Reliene, Alexander J. R. Bishop, Jiri Aubrecht, and Robert H. Schiestl Part III. Recombination as a Tool for Producing Targeted Genetic Modification Gene Targeting at the Chromosomal Immunoglobulin Locus: A Model System for the Study of Mammalian Homologous Recombination Mechanisms Mark D. Baker DNA Fragment Transplacement in Saccharomyces cerevisiae: Some Genetic Considerations Glenn M. Manthey, Michelle S. Navarro, and Adam M. Bailis Targeted Gene Modification Using Triplex-Forming Oligonucleotides Jean Y. Kuan and Peter M. Glazer Using Nucleases to Stimulate Homologous Recombination Dana Carroll Enhancement of In Vivo Targeted Nucleotide Exchange by Nonspecific Carrier DNA Katie K. Maguire and Eric B. Kmiec Part IV. Biochemistry of Recombination Chromatin Immunoprecipitation to Investigate Protein-DNA Interactions During Genetic Recombination Tamara Goldfarb and Eric Alani Holliday Junction Branch Migration and Resolution Assays Angelos Constantinou and Stephen C. West Index

Blurb

Genetic recombination plays an essential role in such vitally important biological processes as DNA repair, regulation of gene expression, disease etiology, meiotic chromosome segregation, and evolution. In Genetic Recombination: Reviews and Protocols, prominent international investigators actively engaged in recombination research describe in great detail their best techniques for studying recombination. The methods range from approaches and model systems to be used in a variety of eukaryotic organisms and in a mammalian parasite, to biochemical analyses useful for furthering the understanding of recombination mechanisms. Additional techniques use recombination as a reporter of genomic instability in lower and higher eukaryotes and as a tool for producing targeted genetic modification. Each readily reproducible method includes step-by-step instructions, a background introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
Cutting-edge and highly practical, Genetic Recombination: Reviews and Protocols offers both novice and experienced investigators an illuminating synthesis of what we currently know about recombination, as well as a wide-ranging collection of powerful tools for elucidating its mechanisms and regulation throughout biology today.