Overview

Nucleases are enzymes that restructure or degrade nucleic acid polymers and are vital to the control of every area of metabolism. Many types of nucleases are used in laboratories and their commercial and clinical uses are expanding, thus this book has been developed to introduce the scientists to cell-characterized protein nucleases, and the methods used to determine their activity, structure interaction with other molecules and their physiological role. Also covered are protocols for studying the inhibitors and activators of nucleases, and methods to determine and model the 3D structure of nucleases and their complexes with substrates and inhibitors. A section covering nucleases in the clinic summarizes assays and protocols suitable for use with tissues and for nuclease-based therapeutics. All the chapters describe why and when to use the assays, and the thinking that went into the development of the protocols. The comments and notes sections provide guidance and insight when things go wrong and how to correct them. The protocols are also easily modifiable to suit the researcher's needs as new developments in the field begin to emerge, thus making this an essential reference for all molecular biologists.

Full Product Details

Author:   Catherine H. Schein
Publisher:   Humana Press Inc.
Imprint:   Humana Press Inc.
Edition:   2001 ed.
Volume:   160
Dimensions:   Width: 17.80cm , Height: 3.00cm , Length: 25.40cm
Weight:   2.760kg
ISBN:  

9780896036796

ISBN 10:   0896036790
Pages:   525
Publication Date:   30 January 2001
Audience:   College/higher education ,  Professional and scholarly ,  Undergraduate ,  Professional & Vocational
Format:   Hardback
Publisher's Status:   Active
Availability:   Out of stock  
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Table of Contents

Characterizing Nuclease Activity.- Processivity of DNA Repair Enzymes.- Analysis by HPLC of Distributive Activities and the Synthetic (Back) Reaction of Pancreatic-Type Ribonucleases.- Ier1 p: A Kinase and Site-Specific Endoribonuclease.- Microtiter-Plate Assay and Related Assays for Nonspecific Endonucleases.- Quantitating mRNAs with Relative and Competitive RT-PCR.- Expressing Self-Incompatibility RNases (S-RNases) in Transgenic Plants.- Molecular Cloning, Tissue Distribution, and Chromosomal Localization of the Human Homolog of the R2/Th/Stylar Ribonuclease Gene Family.- Inhibitors and Activators of Nucleases.- Ribonuclease Inhibitors.- Producing Soluble Recombinant RNases and Assays to Measure Their Interaction with Interferon-? In Vitro?.- Assays for the Evaluation of HIV-1 Integrase Inhibitors.- The Direction of Ribonucleases H by Antisense Oligodeoxynucleotides.- The 2–5A/RNase L Pathway and Inhibition by RNase L Inhibitor (RLI).- Relating Nucleas Structure and Function.- Crystallization and Crystal Structure Determination of Ribonuclease A-Ribonuclease Inhibitor Protein Complex.- Methods for Studying the Interaction of Barnase with Its Inhibitor Barstar.- Assaying In Vitro Refolding of RNases by Mass Spectrometry.- Dissecting Nucleases into Their Structural and Functional Domains.- Using Electrostatics to Define the Active Site of Serratia Endonuclease.- Homology Modeling and Simulations of Nuclease Structures.- Methods for Determining Activity and Specificity of DNA Binding and DNA Cleavage by Class II Restriction Endonucleases.- Engineered Properties and Assays for Human DNase I Mutants.- Nucleases in the Clinic.- Assays for Human DNase I Activity in Biological Matrices.- Evaluation of Retroviral Ribonuclease H Activity.- Assays for Detection of RNase ASuperfamily Ribonucleases.- Assay for Antitumor and Lectin Activity in RNase Homologs.- Isolation and Enzymatic Activity of Angiogenin.- Preparation and Preclinical Characterization of RNase-Based Immunofusion Proteins.- Assays Using Nucleases.- Restriction Endonucleases and Their Uses.- Isolation and Characterization of an Unknown Restriction Endonuclease.- Gene Modification with Hapaxoterministic Restriction Enzymes.- Techniques to Measure Nucleic Acid-Protein Binding and Specificity.- Analyzing the Developmental Expression of Sigma Factors with S1-Nuclease Mapping.- Detection and Quantitation of mRNAs Using Ribonuclease Protection Assays.

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