Nuclease Methods and Protocols

by Catherine H. Schein
Nuclease Methods and Protocols
Author: Catherine H. Schein
Publisher: Springer Science & Business Media
Total Pages: 521
Release: 2008-02-03
Genre: Science
ISBN: 1592592333
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Nucleases, enzymes that restructure or degrade nucleic acid polymers, are vital to the control of every area of metabolism. They range from “housekeeping” enzymes with broad substrate ranges to extremely specific tools (1). Many types of nucleases are used in lab protocols, and their commercial and clinical uses are expanding. The purpose of Nuclease Methods and Protocols is to introduce the reader to some we- characterized protein nucleases, and the methods used to determine their activity, structure, interaction with other molecules, and physiological role. Each chapter begins with a mini-review on a specific nuclease or a nuclease-related theme. Although many chapters cover several topics, they were arbitrarily divided into five parts: Part I, “Characterizing Nuclease Activity,” includes protocols and assays to determine general (processive, distributive) or specific mechanisms. Methods to assay nuclease products, identify cloned nucleases, and determine their physiological role are also included here. Part II, “Inhibitors and Activators of Nucleases,” summarizes assays for measuring the effects of other proteins and small molecules. Many of these inhibitors have clinical relevance. Part III, “Relating Nuclease Structure and Function,” provides an overview of methods to determine or model the 3-D structure of nucleases and their complexes with substrates and inhibitors. A 3-D structure can greatly aid the rational design of nucleases and inhibitors for specific purposes. Part IV, “Nucleases in the Clinic,” summarizes assays and protocols suitable for use with t- sues and for nuclease based therapeutics.

Artificial Nucleases

by Marina A. Zenkova
Artificial Nucleases
Author: Marina A. Zenkova
Publisher: Springer Science & Business Media
Total Pages: 330
Release: 2004-01-07
Genre: Medical
ISBN: 9783540201120
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The development of agents capable of cleaving RNA and DNA has attracted considerable attention from researchers in the last few years, because of the immediate and very important applications they can find in the emerging fields of biotechnology and pharmacology. There are essentially two classes of these agents - nucleases that occur naturally inside cells and synthetically produced artificial nucleases. The first class includes protein enzyme nucle ases and catalytic RNA structured ribozymes that perform cleavage of the phosphodiester bonds in nucleic acids according to a hydrolytic pathway in the course of different biochemical processes in the cell. A different pathway is used by some antibiotics which cleave DNA via redox-based mechanisms resulting in oxidative damage of nucleotide units and breakage of the DNA backbone. The above molecules are indispensable tools for manipulating nucleic acids and processing RNA; DNA-cleaving antibiotics and cytotoxic ribonucleases have demonstrated utility as chemotherapeutic agents. The second class, artificial nucleases, are rationally designed to imitate the active centers of natural enzymes by simple structures possessing minimal sets of the most important characteristics that are essential for catalysis. A dif ferent approach, in vitro selection, was also used to create artificial RNA and DNA enzymes capable of cleaving RNA. Being less efficient and specific as compared to the natural enzymes, the primitive mimics are smaller and robust and can function in a broad range of conditions.

Molecular Biology of Nucleases

by Nawin C. Mishra
Molecular Biology of Nucleases
Author: Nawin C. Mishra
Publisher: CRC Press
Total Pages: 300
Release: 1995-03-06
Genre: Medical
ISBN: 9780849376580
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Nucleases occupy a central position in the biochemistry of DNA transactions and other metabolism of nucleic acids in all organisms. They have also proven useful in modern biological studies crucial for the development of recombinant DNA technology and reverse genetics. Nucleases assist in the identification and characterization of genes responsible for several diseases and their possible alleviation by gene therapy. Molecular Biology of Nucleases introduces the properties and biological roles of nucleases. It is the one comprehensive source for newcomers to the field.

Restriction Endonucleases

by Alfred Pingoud
Restriction Endonucleases
Author: Alfred Pingoud
Publisher: Springer Science & Business Media
Total Pages: 462
Release: 2012-12-06
Genre: Science
ISBN: 3642188516
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Restriction enzymes are highly specific nucleases which occur ubiquitously among prokaryotic organisms, where they serve to protect bacterial cells against foreign DNA. Many different types of restriction enzymes are known, among them multi-subunit enzymes which depend on ATP or GTP hydrolysis for target site location. The best known representatives, the orthodox type II restriction endonucleases, are homodimers which recognize palindromic sequences, 4 to 8 base pairs in length, and cleave the DNA within or immediately adjacent to the recognition site. In addition to their important biological role (up to 10 % of the genomes of prokaryotic organisms code for restriction/modification systems!), they are among the most important enzymes used for the analysis and recombination of DNA. In addition, they are model systems for the study of protein-nucleic acids interactions and, because of their ubiquitous occurence, also for the understanding of the mechanisms of evolution.

Targeted Genome Editing Using Site-Specific Nucleases

by Takashi Yamamoto
Targeted Genome Editing Using Site-Specific Nucleases
Author: Takashi Yamamoto
Publisher: Springer
Total Pages: 206
Release: 2015-01-05
Genre: Science
ISBN: 4431552278
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This book serves as an introduction to targeted genome editing, beginning with the background of this rapidly developing field and methods for generation of engineered nucleases. Applications of genome editing tools are then described in detail, in iPS cells and diverse organisms such as mice, rats, marine invertebrates, fish, frogs, and plants. Tools that are mentioned include zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and CRISPR/Cas9, all of which have received much attention in recent years as breakthrough technologies. Genome editing with engineered nucleases allows us to precisely change the target genome of living cells and is a powerful way to control functional genes. It is feasible in almost all organisms ranging from bacteria to plants and animals, as well as in cultured cells such as ES and iPS cells. Various genome modifications have proven successful, including gene knockout and knock-in experiments with targeting vectors and chromosomal editing. Genome editing technologies hold great promise for the future, for example in biomedical research, clinical medicine, and generation of crops and livestock with desirable traits. A wide range of readers will find this book interesting, and with its focus on applications in a variety of organisms and cells, the book will be valuable for life scientists in all fields.

Diagnostic Molecular Biology

by Chang-Hui Shen
Diagnostic Molecular Biology
Author: Chang-Hui Shen
Publisher: Elsevier
Total Pages: 590
Release: 2023-06-29
Genre: Science
ISBN: 0323986099
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Diagnostic Molecular Biology, Second Edition describes the fundamentals of molecular biology in a clear, concise manner with each technique explained within its conceptual framework and current applications of clinical laboratory techniques comprehensively covered. This targeted approach covers the principles of molecular biology, including basic knowledge of nucleic acids, proteins and chromosomes; the basic techniques and instrumentations commonly used in the field of molecular biology, including detailed procedures and explanations; and the applications of the principles and techniques currently employed in the clinical laboratory. Topics such as whole exome sequencing, whole genome sequencing, RNA-seq, and ChIP-seq round out the discussion. Fully updated, this new edition adds recent advances in the detection of respiratory virus infections in humans, like influenza, RSV, hAdV, hRV but also corona. This book expands the discussion on NGS application and its role in future precision medicine. - Provides explanations on how techniques are used to diagnosis at the molecular level - Explains how to use information technology to communicate and assess results in the lab - Enhances our understanding of fundamental molecular biology and places techniques in context - Places protocols into context with practical applications - Includes extra chapters on respiratory viruses (Corona)

Comparative Virology

by Karl Maramorosch
Comparative Virology
Author: Karl Maramorosch
Publisher: Academic Press
Total Pages: 601
Release: 2014-06-28
Genre: Science
ISBN: 1483269698
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Comparative Virology provides an integrated comparison of viruses, based on their chemical and morphological characteristics. These descriptions will not only give the reader a background but also a detailed analysis of the various groups. In some instances the groups are still host related, as in the case of bacteriophages and polyhedral insect viruses. In others, for instance in pox viruses, the group comprises viruses of vertebrates and invertebrates. The hosts of the bacilliform Rhabdovirales range from man and other warm-blooded vertebrates through invertebrate animals to plants. A special chapter is devoted to viruses devoid of protein—a group that is of great interest and that has only recently been recognized. Since there is historical and practical interest in écologie groupings, such as arboviruses and oncogenic viruses, chapters on such groups have also been included. The book opens with a discussion on the classification of viruses. Chapters dealing with DNA viruses and RNA viruses follow, and the ecologically and disease-oriented groups complete the volume. It is hoped that ""Comparative Virology"" will help bring unity to the science of virology through the comparative approach that is not dependent on virus-host interactions. The combined efforts of eminent contributors to discuss and evaluate new information will hopefully benefit all who are interested in virology

Molecular Anatomy of Cellular Systems

by I. Endo
Molecular Anatomy of Cellular Systems
Author: I. Endo
Publisher: Elsevier
Total Pages: 241
Release: 2002-08-20
Genre: Science
ISBN: 0080536743
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In this book, the progress during the last ten years is reviewed and future plans outlined to realize and establish the concept of design in the biological sciences. Design is a leading concept as well as the principal motivation for the creation of artificial systems. A successful design generally requires that the structures and functions of the elements that constitute the system as well as the principles that determine how the elements cooperate together to create function be fully understood. These requirements have not been satisfied within the fields of biotechnology and medicine. Compared to the recent emergence of artificial systems, living organisms acquired their present day structures and functions through evolution over three to four billion years. Despite the fact that the design of living organisms is recorded in the DNA sequence, our understanding of the structures and functions of the elements that constitute living organisms is very limited. To fulfill the requirements, the following approaches were initiated under a ten-year project entitled "Biodesign Research". Firstly, we tried to isolate and characterize the functional elements that constitute the organelles of various organisms. Secondly, we tried to reconstitute systems that reproduce biological functions in vitro from individual elements in order to understand how the elements cooperate together to yield a function. Thirdly, we attempted to resolve biological structures at various resolutions ranging from the atomic to the cellular level to further our knowledge about the fundamental principles that various functions at the molecular level and to design artificial systems.