Meiosis: Volume 1, Molecular and Genetic Methods

ISBN-13: 9781934115664 / Angielski / Twarda / 2009 / 365 str.

Scott Keeney

Opis

Each generation in a sexually reproducing organism such as a fly or a mouse passes through the bottleneck of meiosis, which is the specialized cell division that gives rise to haploid reproductive cells (sperm, eggs, spores, etc. ). The principal function of meiosis is to reduce the genome complement by half, which is accomplished through sequential execution of one round of DNA replication followed by two rounds of chromosome segregation. Within the extended prophase between DNA replication and the first meiotic division in most organisms, homologous maternal and paternal chromosomes pair with one another and undergo homologous recombination, which establishes physical connections that link the homologous chromosomes until the time they are separated at anaphase I. Recombination also serves to increase genetic diversity from one generation to the next by breaking up linkage groups. The unique chromosome dynamics of meiosis have fascinated scientists for well over a century, but in recent years there has been an explosion of new information about how meiotic chromosomes pair, recombine, and are segregated. Progress has been driven by advances in three main areas: (1) genetic identification of meiosis-defective mutants and cloning of the genes involved; (2) development of direct physical assays for DNA intermediates and products of recombination; and (3) increasingly sophisticated cy- logical methods that describe chromosome behaviors and the spatial and temporal patterns by which specific proteins associate with meiotic chromosomes.

Spis treści

Part I. Genetic Methods for Studying Meiotic Recombination and Chromosome Dynamics Interaction of Genetic and Environmental Factors in Saccharomyces cerevisiae Meiosis: The Devil is in the Details Victoria E. Cotton, Eva R. Hoffmann, Mohammed F. F. Abdullah, and Rhona H. Borts Optimizing Sporulation Conditions for Different Saccharomyces cerevisiae Strain Backgrounds Susan L. Elrod, Sabrina M. Chen, Katja Schwartz, and Elizabeth O. Shuster Modulating and Targeting Meiotic Double-Strand Breaks in Saccharomyces cerevisiae Alain Nicolas Methods for Analysis of Crossover Interference in Saccharomyces cerevisiae Franklin W. Stahl and Elizabeth Housworth Measurement of Spatial Proximity and Accessibility of Chromosomal Loci in Saccharomyces cerevisiae Using Cre/loxP Site-Specific Recombination Doris Lui and Sean M. Burgess Genetic Analysis of Meiotic recombination in Schizosaccharomyces pombe Gerald R. Smith Analysis of Meiotic Recombination in Caenorhabditis elegans Kenneth J. Hillers and Anne M. Villeneuve Visual Markers for Detecting Gene Conversion Directly in the Gametes of Arabidopsis thaliana Luke E. Berchowitz and Gregory P. Copenhaver Part II. Molecular Analysis of Recombination and Protein-DNA Interactions During Meiosis Gel Electrophoresis Assays for Analyzing DNA Double-Strand Breaks in Saccharomyces cerevisiae at Various Spatial Resolutions Hajime Murakami, Valérie Borde, Alain Nicolas, and Scott Keeney Genome-Wide Mapping of Meiotic DNA Double-Strand Breaks in Saccharomyces cerevisiae Cyril Buhler, Robert Shroff, Michael Lichten Detection of Meiotic DNA Breaks in Mouse Testicular Germ Cells Jian Qin, Jaichandar Subramanian, and NormanArnheim End-Labeling and Analysis of Spo11-Oligonucleotide Complexes in Saccharomyces cerevisiae Matthew J. Neale and Scott Keeney Detection of SPO11-Oligonucleotide Complexes from Mouse Testes Jing Pan and Scott Keeney Stabilization and Electrophoretic Analysis of Meiotic Recombination Intermediates in Saccharomyces cerevisiae Steve D. Oh, Lea Jessop, JessicaP. Lao, Thorsten Allers, Michael Lichten, and Neil Hunter Using Schizosaccharomyces pombe Meiosis to Analyze DNA Recombination Intermediates Randy W. Hyppa and Gerald R. Smith Analysis of Chromatin Structure at Meiotic DSB Sites in yeasts Kouji Hirota, Tomoyuki Fukuda, Takatomi Yamada, and Kunihiro Ohta Analysis of Protein-DNA Interactions During Meiosis by Quantitative Chromatin-Immunoprecipitation (qChIP) Marco Antonio Mendoza, Silvia Panizza, and Franz Klein Genome-Wide High-Resolution Chromatin Immunoprecipitation of Meiotic Chromosomal Proteins in Saccharomyces cerevisiae Kazuto Kugou and Kunihiro Ohta Parallel Detection of Crossovers and Non-Crossovers in Mouse Germ Cells Frédéric Baudat and Bernard de Massy Analysis of Meiotic Recombination Products from Human Sperm Liisa Kauppi, Celia A. May, and Alec J. Jeffreys

Blurb

The unique chromosome dynamics of meiosis have fascinated scientists for well over a century, but in recent years there has been a proliferation of new data concerning the pairing, recombination, and segregation of chromosomes. In Meiosis, expert researchers explore recent advances in three main areas: quantitative genetic methods for analysis of homologous recombination and chromosome pairing, development of direct physical assays for DNA intermediates and products of recombination, and cytological methods for characterizing chromosome behaviors and the patterns by which specific proteins associate with meiotic chromosomes. Meiosis: Volume 1, Molecular and Genetic Methods details methods for culturing and manipulating commonly used model organisms and protocols for detecting and quantifying meiotic recombination events through genetic and physical analysis of DNA-DNA or Protein-DNA interactions. Methods are provided for studies in yeasts, plants, and animals, including mouse and human. Composed in the highly successful Methods in Molecular Biology(tm) series format, each chapter contains a brief introduction, step-by-step methods, a list of necessary materials, and a Notes section which shares tips on troubleshooting and avoiding known pitfalls.

Comprehensive and cutting-edge, Meiosis: Volume 1, Molecular and Genetic Methods is an essential guidebook, providing up-to-date and critical new protocols for the study of meiotic chromosome dynamics.