Genetic Recombination

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.

Includes supplementary material: sn.pub/extras

Klappentext
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.

Inhalt
Studying Recombination Events in Eukaryotes.- Determination of Mitotic Recombination Rates by Fluctuation Analysis in Saccharomyces cerevisiae.- Determination of Intrachromosomal Recombination Rates in Cultured Mammalian Cells.- Intrachromosomal Homologous Recombination in Arabidopsis thaliana.- Analysis of Recombinational Repair of DNA Double-Strand Breaks in Mammalian Cells With I-SceI Nuclease.- Transformation of Monomorphic and Pleomorphic Trypanosoma brucei.- Forward Genetic Screens for Meiotic and Mitotic Recombination-Defective Mutants in Mice.- Recombination as a Reporter of Genomic Instability.- Detecting Carcinogens With the Yeast DEL Assay.- In Vivo DNA Deletion Assay to Detect Environmental and Genetic Predisposition to Cancer.- Recombination as a Tool for Producing Targeted Genetic Modification.- Gene Targeting at the Chromosomal Immunoglobulin Locus.- DNA Fragment Transplacement in Saccharomyces cerevisiae.- Targeted Gene Modification Using Triplex-Forming Oligonucleotides.- Using Nucleases to Stimulate Homologous Recombination.- Enhancement of In Vivo Targeted Nucleotide Exchange by Nonspecific Carrier DNA.- Biochemistry of Recombination.- Chromatin Immunoprecipitation to Investigate Protein-DNA Interactions During Genetic Recombination.- Holliday Junction Branch Migration and Resolution Assays.