Download or read book In Vitro Mutagenesis written by Andrew Reeves and published by Humana. This book was released on 2016-10-06 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: In vitro mutagenesis remains a critical experimental approach for investigating gene and protein function at the cellular level. This volume provides a wide variety of updated and novel approaches for performing in vitro mutagenesis using such methods as genome editing, transposon (Tn) mutagenesis, site-directed, and random mutagenesis. In Vitro Mutagenesis: Methods and Protocols guides readers through methods for gene and genome editing, practical bioinformatics approaches for identifying mutagenesis targets, and novel site-directed and random mutagenesis approaches aimed at gaining a better understanding of protein-protein and protein-cofactor interactions. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, In Vitro Mutagenesis: Methods and Protocols aims to provide a highly accessible and practical manual for current and future molecular biology researchers, from the beginner practitioner to the advanced investigator in fields such as molecular genetics, biochemistry, and biochemical and metabolic engineering.
Download or read book In Vitro Mutagenesis Protocols written by Jeff Braman and published by Springer Science & Business Media. This book was released on 2008-02-05 with total page 284 pages. Available in PDF, EPUB and Kindle. Book excerpt: Hands-on researchers with proven track records describe in stepwise fashion their advanced mutagenesis techniques. The contributors focus on improvements to conventional site-directed mutagenesis, including a chapter on chemical site-directed mutagenesis, PCR-based mutagenesis and the modifications that allow high throughput mutagenesis experiments, and mutagenesis based on gene disruption (both in vitro- and in situ-based). Additional methods are provided for in vitro gene evolution; for gene disruption based on recombination, transposon, and casette mutagenesis; and for facilitating the introduction of multiple mutations. Time-tested and highly practical, the protocols in In Vitro Mutagenesis Protocols, 2nd Edition offer today's molecular biologists reliable and powerful techniques with which to illuminate the proteome.
Download or read book Yeast Protocols written by Ivor Howell Evans and published by Springer. This book was released on 1996 with total page 433 pages. Available in PDF, EPUB and Kindle. Book excerpt: Yeast Protocols contains many key techniques for studying the biology of yeasts at both the cellular and molecular levels. Working primarily from Saccharomyces cerevisiae, the expert contributors explain step-by-step how to successfully isolate, identify, and culture yeasts; the secrets of meiotic mapping; how to use PFGE in karyotyping and gene localization; the methods for purification and analysis of various cell components; and the construction and exploitation of genomic DNA clone banks. They also cover the latest methods for chromosome engineering, insertional mutagenesis by Ty elements, mRNA abundance and half-life measurements, the use of reporter gene systems, genotoxicity testing, and more. Yeast Protocols follows the widely applauded Humana Methods in Molecular Biology style: brief introductions putting the particular method in context, comprehensive lists of materials, cookbook style instructions, and troubleshooting notes to avoid common pitfalls and solve problems. The techniques can be used with confidence and success by both inexperienced newcomers and established researchers.
Download or read book Molecular Embryology written by Paul T. Sharpe and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 752 pages. Available in PDF, EPUB and Kindle. Book excerpt: Most people have some interest in embryos; this probably results, in part, from their interest in understanding the biological origins of themselves and their offspring and, increasingly, concerns about how environmental change such as pollution might affect human development. Obviously, et- cal considerations preclude experimental studies of human embryos and, c- sequently, the developmental biologist has turned to other species to examine this process. Fortunately, the most significant conclusion to be drawn from the experimental embryology of the last two decades is the manner in which orthologous or closely related molecules are deployed to mediate similar - velopmental processes in both vertebrates and invertebrates. The molecular mechanisms regulating processes fundamental to most animals, such as axial patterning or axon guidance, are frequently conserved during evolution. (It is now widely believed that the differences between phyla and classes are the result of new genes, arising mostly by duplication and divergence of extant sequences, regulating the appearance of derived characters. ) Other vertebrates are obviously most likely to use the same devel- mental mechanisms as humans and, within the vertebrate subphylum, the - parent degree of conservation of developmental mechanism is considerable. It has long been recognized that particular vertebrate species offer either d- tinct advantages in investigating particular stages of development or are - pecially amenable to particular manipulations. No single animal can provide all the answers because not all types of experiments can be carried out on a single species.
Download or read book Oligodendrocytes written by David A. Lyons and published by Humana Press. This book was released on 2019-03-01 with total page 411 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume looks at the study of oligodendrocytes through in vitro and in vivo techniques, multiple model organisms, using approaches that bridge scales from molecular through system. Chapters in this book cover topics such as fundamental molecular analyses of oligodendrocytes and myelin; in vitro, ex vivo, and in vivo molecular-cellular-electrophysiology-based techniques; oligodendrocyte formation, homeostasis, and disruption in zebrafish and Xenopus; and parallel system-level imaging of animal and human models. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and thorough, Oligodendrocytes: Methods and Protocols is a valuable reference guide that highlights the expansive and fast-paced nature of research into oligodendrocyte biology underlying health and function.
Download or read book Directed Evolution Library Creation written by Frances H. Arnold and published by Humana Press. This book was released on 2010-11-10 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Biological systems are very special substrates for engineering—uniquely the products of evolution, they are easily redesigned by similar approaches. A simple algorithm of iterative cycles of diversification and selection, evolution works at all scales, from single molecules to whole ecosystems. In the little more than a decade since the first reported applications of evolutionary design to enzyme engineering, directed evolution has matured to the point where it now represents the centerpiece of industrial biocatalyst development and is being practiced by thousands of academic and industrial scientists in com- nies and universities around the world. The appeal of directed evolution is easy to understand: it is conceptually straightforward, it can be practiced without any special instrumentation and, most important, it frequently yields useful solutions, many of which are totally unanticipated. Directed evolution has r- dered protein engineering readily accessible to a broad audience of scientists and engineers who wish to tailor a myriad of protein properties, including th- mal and solvent stability, enzyme selectivity, specific activity, protease s- ceptibility, allosteric control of protein function, ligand binding, transcriptional activation, and solubility. Furthermore, the range of applications has expanded to the engineering of more complex functions such as those performed by m- tiple proteins acting in concert (in biosynthetic pathways) or as part of mac- molecular complexes and biological networks.
Download or read book Somaclonal Variation and Induced Mutations in Crop Improvement written by S.M. Jain and published by Springer Science & Business Media. This book was released on 2013-03-14 with total page 615 pages. Available in PDF, EPUB and Kindle. Book excerpt: Genetic variability is an important parameter for plant breeders in any con ventional crop improvement programme. Very often the desired variation is un available in the right combination, or simply does not exist at all. However, plant breeders have successfully recombined the desired genes from cultivated crop gerrnplasm and related wild species by sexual hybridization, and have been able to develop new cultivars with desirable agronomie traits, such as high yield, disease, pest, and drought resistance. So far, conventional breeding methods have managed to feed the world's ever-growing population. Continued population growth, no further scope of expanding arable land, soil degradation, environ mental pollution and global warrning are causes of concern to plant biologists and planners. Plant breeders are under continuous pressure to improve and develop new cultivars for sustainable food production. However, it takes several years to develop a new cultivar. Therefore, they have to look for new technologies, which could be combined with conventional methods to create more genetic variability, and reduce the time in developing new cultivars, with early-maturity, and improved yield. The first report on induced mutation of a gene by HJ. Muller in 1927 was a major mi1estone in enhancing variation, and also indicated the potential applica tions of mutagenesis in plant improvement. Radiation sources, such as X-rays, gamma rays and fast neutrons, and chemical mutagens (e. g. , ethyl methane sulphonate) have been widely used to induce mutations.
Download or read book Directed Enzyme Evolution written by Frances H. Arnold and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 381 pages. Available in PDF, EPUB and Kindle. Book excerpt: Directed evolution comprises two distinct steps that are typically applied in an iterative fashion: (1) generating molecular diversity and (2) finding among the ensemble of mutant sequences those proteins that perform the desired fu- tion according to the specified criteria. In many ways, the second step is the most challenging. No matter how cleverly designed or diverse the starting library, without an effective screening strategy the ability to isolate useful clones is severely diminished. The best screens are (1) high throughput, to increase the likelihood that useful clones will be found; (2) sufficiently sen- tive (i. e. , good signal to noise) to allow the isolation of lower activity clones early in evolution; (3) sufficiently reproducible to allow one to find small improvements; (4) robust, which means that the signal afforded by active clones is not dependent on difficult-to-control environmental variables; and, most importantly, (5) sensitive to the desired function. Regarding this last point, almost anyone who has attempted a directed evolution experiment has learned firsthand the truth of the dictum “you get what you screen for. ” The protocols in Directed Enzyme Evolution describe a series of detailed p- cedures of proven utility for directed evolution purposes. The volume begins with several selection strategies for enzyme evolution and continues with assay methods that can be used to screen enzyme libraries. Genetic selections offer the advantage that functional proteins can be isolated from very large libraries s- ply by growing a population of cells under selective conditions.
Download or read book Manual on MUTATION BREEDING THIRD EDITION written by Food and Agriculture Organization of the United Nations and published by Food & Agriculture Org.. This book was released on 2018-10-09 with total page 319 pages. Available in PDF, EPUB and Kindle. Book excerpt: This paper provides guidelines for new high-throughput screening methods – both phenotypic and genotypic – to enable the detection of rare mutant traits, and reviews techniques for increasing the efficiency of crop mutation breeding.
Download or read book Safety of Genetically Engineered Foods written by National Research Council and published by National Academies Press. This book was released on 2004-07-08 with total page 254 pages. Available in PDF, EPUB and Kindle. Book excerpt: Assists policymakers in evaluating the appropriate scientific methods for detecting unintended changes in food and assessing the potential for adverse health effects from genetically modified products. In this book, the committee recommended that greater scrutiny should be given to foods containing new compounds or unusual amounts of naturally occurring substances, regardless of the method used to create them. The book offers a framework to guide federal agencies in selecting the route of safety assessment. It identifies and recommends several pre- and post-market approaches to guide the assessment of unintended compositional changes that could result from genetically modified foods and research avenues to fill the knowledge gaps.
Download or read book Gene Knockout Protocols written by Ralf Kühn and published by Humana Press. This book was released on 2009-03-27 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Following the completion of the mouse and human genome sequences, a major challengeisthefunctionalcharacterizationofeverymammaliangeneandthedeciph- ing of their molecular interaction network. The mouse offers many advantages for the use of genetics to study human biology and disease, unmatched among other m- mals. Its development, body plan, physiology, behavior, and diseases have much in common, based on the fact that 99% of the human genes have a mouse ortholog. The investigation of gene function using mouse models is based on many years of tech- logical development. In the two decades since gene targeting in murine embryonic stem (ES) cells was first described by Mario Capecchi and colleagues, more than 3000 predesigned mouse mutants have been developed. To date, a variety of mouse mutagenesis techniques, either gene- or phenotype-driven, are used as systematic approaches. The availability of the genome sequence supports gene-driven approaches such as gene-trap and targeted mutagenesis in ES cells, allowing efficient and precise gene disruption. In combination with the use of site-specific DNA recombinases, in particular the Cre/loxP system, gene disruptioncan be directed to specific cell types in conditionalmousemutants. Furthermore,chemicalandtransposonmutagenesisofthe mouse genome enables us to perform phenotype-driven screens for the unbiased identification of phenotype–genotype correlations involved in models of human d- ease. Over the next several years, the mouse genome will be systematically altered, and the techniques for achieving predesigned manipulations will be constantly developed further and improved. The second edition of Gene Knockout Protocols brings together distinguished c- tributorswithextensiveexperienceinthegenetargetingandmousegeneticsfields.
Download or read book PCR Cloning Protocols written by Bing-Yuan Chen and published by Humana. This book was released on 2010-11-10 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: PCR Cloning Protocols, Second Edition, updates and expands Bruce White's best-selling PCR Cloning Protocols (1997) with the newest procedures for DNA cloning and mutagenesis. Here the researcher will find readily reproducible methods for all the major aspects of PCR use, including PCR optimization, computer programs for PCR primer design and analysis, and novel variations for cloning genes of special characteristics or origin, with emphasis on long distance PCR and GC-rich template amplification. Also included are both conventional and novel enzyme-free and restriction site-free procedures to clone PCR products into a range of vectors, as well as state-of-the-art protocols to facilitate DNA mutagenesis and recombination, and to clone the challenging uncharacterized DNA flanking a known DNA fragment.
Download or read book Molecular Biology of the Cell written by and published by . This book was released on 2002 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book B Cell Protocols written by Hua Gu and published by Springer Science & Business Media. This book was released on 2008-02-04 with total page 294 pages. Available in PDF, EPUB and Kindle. Book excerpt: B-lymphocyte development and function remains an exciting area of research for those interested in the physiology and pathology of the immune system in higher animals. While recent advances in genetics and cellular and molecular biology have provided a large spectrum of powerful new experimental tools in this field, it is both time consuming and often very difficult for a student or just any bench-side worker to identify a reliable experimental protocol in the ocean of the literature. The aim of B Cell Protocols is to provide a collection of diverse protocols ranging from the latest inventions and applications to some classic, but still frequently used methods in B-cell biology. The authors of the various chapters are all highly qualified scientists who are either the inventors or expert users of these methods. Their extensive experience in mastering a particular method provides not only the step-by-step details of a reproducible protocol, but also useful troubleshooting tips that readers will appreciate in their daily work. We hope that this book will be helpful for both beginning and experienced researchers in the field in designing or modifying an experimental approach, and exploring a biological question from multiple angles.
Download or read book TET Proteins and DNA Demethylation written by Ozren Bogdanovic and published by Humana. This book was released on 2021-05-20 with total page 320 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume explores the latest methods used to study various aspects of TET proteins and their biology. Chapters in this book are divided into five parts. Part One describes technologies aimed at detecting and quantifying DNA methylation turnover using massively parallel sequencing, ELISA, and mass spectrometry approaches. Part Two looks at data analyses protocols for distinguishing acting versus passive DNA demethylation and estimation of 5mC and 5hmC levels. Part Three deals with a new topic that takes advantage of modified CRISPR/Cas9 genome editing systems to target DNA demethylation activity to genomic loci of interest. Part Four discusses protocols that detail how to purify TET proteins and unravel their protein interactions, and Part Five looks at the assessment of TET protein function and activity in vivo and in vitro. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and thorough, TET Proteins and DNA Demethylation: Methods and Protocols is a valuable resource that aims to help research scientists at all levels working in the fields of DNA demethylation dynamics. Chapters 3, 7 and 17 are available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.
Download or read book PCR Cloning Protocols written by Bruce Alan White and published by . This book was released on 1997 with total page 490 pages. Available in PDF, EPUB and Kindle. Book excerpt: Distinguished scientists and researchers present a comprehensive collection of current preparative PCR techniques that can be used in cloning and modifying DNA and cDNA. Topics include performing and optimizing PCR (including long PCR), cloning PCR products, cloning unknown neighboring DNA, and library construction and screening. Also covered are mutagenesis, recombination, and in vitro selection, differential and subtractive approaches to cDNA analysis and screening, and cloning members of gene families. The techniques bring to both new and established researchers the power to apply PCR-based methodology to the cloning and modification of DNA, either through innovative protocols or by fostering individual creativity to modify and customize the protocols to best fit their own needs.
Download or read book Peptide and Protein Drug Analysis written by Ronald Reid and published by CRC Press. This book was released on 1999-11-12 with total page 918 pages. Available in PDF, EPUB and Kindle. Book excerpt: Furthering efforts to simulate the potency and specificity exhibited by peptides and proteins in healthy cells, this remarkable reference supplies pharmaceutical scientists with a wealth of techniques for tapping the enormous therapeutic potential of these molecules-providing a solid basis of knowledge for new drug design. Provides a broad, comprehensive overview of peptides and proteins as mediators of cell movement, proliferation, differentiation, and communication. Written by more than 50 leading international authorities, Peptides and Protein Drug Analysis discusses strategies for dealing with the complexity of peptides and proteins in conformational flexibility and amino acid sequence variability analyzes drug formulations facilitated by solid-phase peptide synthesis and recombinant DNA technology examines chemical purity analysis by high-pressure chromatographic, capillary electrophoretic, gel electrophoretic, and isoelectric focusing methods highlights drug design elements derived from protein folding, bioinformatics, and computational chemistry demonstrates uses of unnatural mutagenesis and combinatorial chemistry explores mass spectrometry, protein sequence, and carbohydrate analysis illustrates bioassays and other new functional analysis methods surveys spectroscopic techniques such as ultraviolet, fluorescence, Fourier transform infrared, and nuclear magnetic resonance (NMR) addresses ways of distinguishing between levels of therapeutic and endogenous agents in cells reviews structural analysis tools such as ultracentrifugation and light, X-ray, and neutron scattering and more! Featuring over 3400 bibliographic citations and more than 500 tables, equations, and illustrations, Peptide and Protein Drug Analysis is a must-read resource for pharmacists; pharmacologists; analytical, organic, and pharmaceutical chemists; cell and molecular biologists; biochemists; and upper-level undergraduate and graduate students in these disciplines.
