Title Nuclease methods and protocols / edited by Catherine H. Schein

Published Totowa, NJ : Humana Press, ©2001

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Description 1 online resource (xvii, 525 pages) : illustrations (some color) Series Methods in molecular biology ; v. 160 Methods in molecular biology (Clifton, N.J.) ; v. 160 Contents Part 1 -- Characterizing nuclease activity -- Part II -- Inhibitors and activators of nucleases -- Part III -- Relating nuclease structure and function -- Part IV -- Nucleases in the clinic -- Part V -- Assays using nucleases Processivity of DNA repair enzymes / R.S. Lloyd -- Analysis by hplc of distributive activities and the synthetic (back) reaction of pancreatic-type ribonucleases / M.V. Nogues and C.M. Cuchillo -- Ire1p: A kinase and site-specific endoribonuclease / T.N. Gonzalez and P. Walter -- Microtiter-plate assay and related assays for nonspecific endonucleases / G. Meiss [and others] -- Quantitating mrnas with relative and competitive RT-CPR / E.A. Prediger -- Expressing self-incompatibility RNases (s-RNases) in transgenic plants / B. Beecher and B.A. McClure -- Molecular cloning, tissue distribution, and chromosomal localization of the human homolog of the R2/Th/Stylar ribonuclease gene family / F. Acquati [and others] -- Ribonuclease inhibitors / B.L. Pasloske -- Producing soluble recombinant RNases and assays to measure their interaction with interferon-[gamma] in vitro / C.H. Schein -- Assays for the evaluation of HIV-1 integrase inhibitors / Z. Debyser [and others] -- The direction of ribonucleases H by antisense oligodeoxynucleotides / R.V. Giles and D.M. Tidd -- The 2-5a/RNase l pathway and inhibition by RNase l inhibitor (rli) / C. Bisbal [and others] -- Crystallization and crystal structure determination of ribonuclease A-ribonuclease inhibitor protein complex / B. Kobe -- Methods for studying the interaction of baRNase with its inhibitor barstar / G. Schreiber -- Assaying in vitro refolding of RNases by mass spectrometry / G. Marino, P. Pucci and M. Ruoppolo -- Dissecting nucleases into their structural and functional domains. Mapping the rna-binding surface of RNase III by NMR / A. Ramos and A. Pastore -- Using electrostatics to define the active site of serratia endonuclease / K.L. Krause and M.D. Miller -- Homology modeling and simulations of nuclease structures / K.V. Soman [and others] -- Methods for determining activity and specificity of DNA binding and DNA cleavage by class II restriction endonucleases / A. Jeltsch and A.M. Pingoud -- Engineered properties and assays for human DNAse i mutants / C.Q. Pan, D.V. Sinicropi and R.A. Lazarus -- Assays for human DNAse i activity in biological matrices / D.V. Sinicropi and R.A. Lazarus -- Evaluation of retroviral ribonuclease h activity / J.T. Miller, J.W. Rausch and S.F. Le Grice -- Assays for detection of RNase a superfamily ribonucleases / H.F. Rosenberg and J.B. Domachowske -- Assay for antitumor and lectin activity in RNase homologs / K. Nitta -- Isolation and enzymatic activity of angiogenin / J.F. Riordan and R. Shapiro -- Preparation and preclinical characterization of RNase-based immunofusion proteins / D.L. Newton and S.M. Rybak -- Restriction endonucleases and their uses / R.J. Williams -- Isolation and characterization of an unknown restriction endonuclease / R.J. Williams -- Gene modification with hapaxoterministic restriction enzymes. Easing the way / S.L. Berger -- Techniques to measure nucleic acid-protein binding and specificity. Nuclear extract preparations, DNAse i footprinting, and mobility shift assays / R.A. Rippe, D.A. Brenner and A. Tugores -- Analyzing the developmental expression of sigma factors with S1-nuclease mapping / J. Kormanec -- Detection and quantitation of mRNAs using ribonuclease protection assays / E.A. Prediger Summary Nucleases, the enzymes that cleave and shape nucleic acids, are key regulators of cellular metabolism. Angiogenesis and the unfolded protein response are just a few instances of important intra- and extracellular pathways regulated by nucleases. In Nuclease Methods and Protocols, researchers at the forefront of academic research and pharmaceutical drug development from four continents summarize their recent results. Their projects involve nucleases in many different areas of medicine and biology, ranging from protein structure and folding, to DNA repair, to developing cures for insidious diseases such as cystic fibrosis, AIDS, and cancer. Each chapter contains a minireview of a specific nuclease or nuclease-related theme, a discussion of why and when to use the assays described, and often, an explanation of how the assay was developed. The authors describe their most valuable methods for determining the mode of action, structure, interaction with other molecules, and physiological role of nucleases. Detailed step-by-step instructions and notes on avoiding pitfalls help ensure readily reproducible results. Comprehensive and highly practical, Nuclease Methods and Protocols offers both novice and experienced researchers a deeper understanding of the importance of nucleases in cell metabolism, growth control mechanisms, and, increasingly, in diagnostics and therapeutics Bibliography Includes bibliographical references and index Notes English In Springer Protocols Subject Nucleases -- Laboratory manuals Ribonucleases. Endonucleases. Deoxyribonucleases Ribonucleases Endonucleases SCIENCE -- Life Sciences -- Biochemistry. Ribonucleases Endonucleases Nucleases Genre/Form Laboratory manuals Laboratory manuals. Manuels de laboratoire. Form Electronic book Author Schein, Catherine H ISBN 9781592592333 0896036790 9780896036796 0585403287 9780585403281 1592592333 1280830360 9781280830365 9786610830367 6610830363

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