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DNA Repair Protocols

Eukaryotic Systems

Series: Methods in Molecular Biology

by Daryl S. Henderson

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Main description:

The field of eukaryotic DNA repair is enjoying a period of remarkable growth and discovery, fueled by technological advances in molecular biol- ogy, protein biochemistry, and genetics. Notahle achievements include the molecular cloning of multiple genes associated with classical human repair disorders, such as xeroderma pigmentosum, Cockayne syndrome, and ataxia telangiectasia; elucidation of the core reaction of nucleotide excision repair (NER); the discovery that certain NER proteins participate not only in repair, but also in transcription; recognition of the crucial role played by mismatch repair processes in maintenance of genome stability and avoidance of cancer; the findings that the tumor suppressor protein p53 is mutated in many types of cancer, and has a key role in directing potentially malignant, genotoxin-dam- aged cells towards an apoptotic fate; and the discovery and elaboration of DNA darnage (and replication) checkpoints, which placed repair phenomenol- ogy firmly within a cell-cycle context. Of course, much remains to be learned about DNA repair.
Tothat end, DNA Repair Protocols: Eukaryotic Systems is about the tools and techniques that have helped propel the DNA repair field into the mainstream of biological research. DNA Repair Protoco/s: Eukaryotic Systems provides detailed, step-by- step instructions for studying manifold aspects of the eukaryotic response to genomic injury. The majority of chapters describe methods for analyzing DNA repair processes in mammalian cells. However, many ofthose techniques can be applied with only minor modification to other systems, and vice versa.

Contents:

Part I. Mutant Isolation and Gene Cloning. Isolation of DNA Structure-dependent Checkpoint Mutants in S. pombe, Rui G. Martinho and Antony M. Carr. Isolating Mutants of the Nematode Caenorhabditis elegans that are Hypersensitive to DNA-damaging Agents, Phil S. Hartman and Naoaki Ishii. Isolating DNA Repair Mutants of Drosophila melanogaster, Daryl S. Henderson. Generation, Identification, and Characterization of Repair-defective Mutants of Arabidopsis, Anne Britt and Cai-Zhong Jiang. Screening for y-ray Hypersensitive Mutants of Arabidopsis, Corinne Davies. Isolation of Mutagen-sensitive Chinese Hamster Cell Lines by Replica Plating, Margaret Zdzienicka. Strategies for Cloning Mammalian DNA Repair Genes, Larry H. Thompson. Novel Complementation Assays for DNA Repair Deficient Cells: Transient and Stable Expression of DNA Repair Genes, Lin Zeng, Alain Sarasin and Mauro Mezzina. Part II. Recognition and Removal of Inappropriate or Damaged DNA Bases. The Use of Electrophoretic Mobility Shift Assays to Study DNA Repair, Byung Joon Hwang, Vaughn Smider and Gilbert Chu. Mismatch Repair Assay, Stephanie Corrette-Bennett/ Robert S. Lahue. Analysis of Cyclobutane-pyrimidine-dimer and (6-4) Photoproduct Photolyases in Crude Extracts: Chromatographic and PCR-amplifiability assays, John Hays and Peter Hoffman. A Dot Blot Immunoassay for UV Photoproducts, Shirley McCready. Measurement of Ultraviolet Radiation-induced DNA Damage Using Specific Antibodies, Ann Stapleton. Quantification of Photoproducts in Mammalian Cell DNA Using Radioimmunoassay, David Mitchell. UV-endonuclease/ alkaline-sedimentation assay for Cyclobutane Pyrimidine Dimers in DNA from UV-irradiated Arabidopsis Plants, John B. Hays and Quishen Pang. DNA Damage Quantitation by Alkaline Gel Electrophoresis, Betsy M. Sutherland, Paula V. Bennett and John C. Sutherland. The Single Cell Gel Test (Comet Assay): A Sensitive Genotoxicity Test for the Detection of DNA Damage and Repair, Gunter Speit and AndreasHartmann. Measuring the Formation and Repair of UV Photoproducts by Ligation-mediated PCR, Gerd P. Pfeifer and Reinhard Dammann. PCR-based Repair Assay I., Keith Grimaldi. PCR-based Repair Assay II.,Keith Grimaldi. Gene-specific and Mitochondrial Repair of Oxidative DNA Damage, Michael R. Ansan and Vilhelm A. Bohr. Characterization of DNA Strand Cleavage by Enzymes That Act at Abasic Sites in DNA, Walter Deutsch and Adly Yacoub. Base Excision repair Assay Using Xenopus laevis Oocyte Extracts, Yoshihiro Matsumoto. In Vitro Base Excision Repair Assay Using Mammalian Cell Extracts, Guido Frosina, Enrico Cappelli, Paola Fortini, Eugenia Dogliotti. Nucleotide Excision Repair in Saccharomyces cerevisiae Whole Cell Extracts, Johnson M.S. Wong, Zhigang He and C. James Ingles. In Vitro Excision Repair Assay in Schizosaccharomyces pombe, Bernard Salles and Patrick Calsou. In Vitro Nucleotide Excision Repair Assay Using Drosophila Cell Lines, Kenji Kohno. Nucleotide Excision Repair in Nuclear Extracts from Xenopus oocytes, Eric J. Ackerman, Lilian K. Koriazova, Jitendra K. Saxena and Alexander Y. Spoonde. Assay for Nucleotide Excision Repair Protein Activity Using Fractionated Cell Extracts and UV-damaged Plasmid DNA, Maureen Biggerstaff and Richard D. Wood. Dual Incision Assays for Nucleotide Excision Repair Using DNA with a Lesion at a Specific Site. Mahmud K.K. Shivji, Jonathan G. Moggs, Isao Kuraoka and Richard D. Wood. In Vitro Chemiluminescence Assay to Measure Excision Repair in Cell Extracts, Bernard Salles and Christian Provot. Part III. DNA Strand Breakage and Repair. Physical Monitoring of HO-induced Homologous Recombination. Allyson Holmes and James E. Haber. Use of P-element Transposons to Generate Site-specific DNA Double-strand Breaks in Drosophila, Satnam S. Banga and Daryl S. Henderson. Analyzing Double-strand Repair Events in Drosophila melanogaster. Gregory Gloor, Tammy Dray and Kathy Keeler. Expression of I-Sce I in Drosophila to Induce DNA Double-stra

PRODUCT DETAILS

ISBN-13: 9781468471441
Publisher: Springer (Humana Press Inc.)
Publication date: March, 2013
Pages: 641
Weight: 964g
Availability: Available
Subcategories: Biochemistry

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