This manual is designed as an intensive introduction to the various tools of molecular biology. It introduces all the basic methods of molecular biology including cloning, PCR, Southern (DNA) blotting, Northern (RNA) blotting, Western blotting, DNA sequencing, oligo-directed mutagenesis, and protein expression.
Key Features
* Provides well-tested experimental protocols for each technique
* Lists the reagents and preparation of each experiment separately
* Contains a complete schedule of experiments and the preparation required
* Includes study questions at the end of each chapter

Nitric Oxide and Other Small Signalling Molecules, Volume 72, the latest release in the Advances in Microbial Physiology series, continues the long tradition of topical, important, cutting-edge reviews in microbiology. The book contains updates in the field, with comprehensive chapters covering the Biochemistry of cysteine persulfides, NO signaling in yeast, The Inflammasome: Regulation of Nitric Oxide and Antimicrobial Host Defense, Nitric Oxide, aN Old molecule with NOble functions in Pseudomonas aeruginosa biology, the Emerging roles of nitric oxide synthase in bacterial physiology, and Anaerobic bacterial response to nitrosative stress.

Bifidobacteria are Gram-positive anaerobic bacteria, found naturally in the gut of humans and other mammals. They are widely used as probiotic organisms in a vast array of formulations for the prevention, alleviation, and treatment of many intestinal disorders. However, bifidobacteria are fastidious microorganisms and are difficult to study in the laboratory, so until recently, understanding of their genetics lagged behind that of other high GC content Gram-positive bacteria. The application of modern whole genome approaches to bifidobacteria research has changed all of this, permitting the accumulation of an impressive amount of data, something that could not have been foreseen a few years ago. This book brings together the expertise and enthusiasm of the leading bifidobacteria researchers from around the world to provide a state-of-the art overview of the molecular biology and genomics of this exciting and important microbial genus. Topics include: ecology, genomics, comparative geno

The Evolution of Molecular Biology: The Search for the Secrets of Life provides the historical knowledge behind techniques founded in molecular biology, also presenting an appreciation of how, and by whom, these discoveries were made. It deals with the evolution of intellectual concepts in the context of active research in an approachable language that accommodates readers from a variety of backgrounds. Each chapter contains a prologue and epilogue to create continuity and provide a complete framework of molecular biology. This foundational work also functions as a historical and conceptual supplement to many related courses in biochemistry, biology, chemistry, genetics and history of science. In addition, the book demonstrates how the roots of discovery and advances-and an individual's own research-have grown out of the history of the field, presenting a more complete understanding and context for scientific discovery.

Metabolic Aspects of Aging, Volume 155, the latest release in the Progress in Molecular Biology and Translational Science series seeks to provide the most topical, informative and exciting monographs available on a wide variety of research topics related to prions, viruses, bacteria and eukaryotes. The series includes in-depth knowledge on the molecular, biological aspects of organismal physiology and function, with this release including chapters on Longevity, Metabolic Disease and Community Health, the Metabolic Aspects of Aging, Obesity, Metabolism, and Aging: A Multiscalar Approach, The Intersection of Curandismo and Western Medicine and Their Epidemiological Impact for Aging Mexican Americans, and more.

The inflammasome is a protein complex composed of an intracellular sensor-typically a Nod-like receptor (NLR), the precursor procaspase-1, and the adaptor ASC. Inflammasome activation leads to the maturation of caspase-1 and the processing of its substrates, IL-1 and IL-18. The inflammasome has been implicated numerous diseases, and blockade of inflammasome-derived IL-1 has beneficial effects on several of these diseases. Different books have been edited about the biology of inflammasomes and about methods to study, however, the implication of this complex in the different diseases and pathological conditions show the need of a book about the clinical implications and therapeutic options. This project will show the context where inflammasomes are being studied and the molecular implications in the medical and clinical contexts. Other important topic of the inflammasomes will be the development of pharmacological inhibitors in order to improve new clinical applications. In this sense, we can find new drugs with inhibitory effects or old drugs with an inhibitory potential effect. There is a need for re-establishing the real benefits of the inflammasome inhibitions in pathological situations and the management of the differents diseases where inflammasomes are implicated.

From small beginnings in the early 1970s, the study of complement regulatory proteins has grown in the last decade to the point where it dominates the complement field. This growth has been fueled by the discovery of new regulators, the cloning of old and new regulators, the discovery that many of the regulators are structurally and evolutionarily related to each other and the development of recombinant forms for use in therapy. There are now more proteins known to be involved in controlling the complement system than there are components of the system and the list continues to grow. The time is ripe for a comprehensive review of our current knowledge of these intriguing proteins. This book does just that. The first few chapters discuss the "nuts-and-bolts" of the complement regulators, describing their structures, functional roles and modes of action. The roles of the complement regulators "in vivo" are then described, focusing on the consequences of deficiency, roles in the reproductive system, interactions with pathogens and exploitation for therapy. The interesting developments in defining the complement regulators expressed in other species are also discussed. The book is written as a monograph, albeit by two people. The text is as readable as possible without compromising on scientific accuracy and completeness. The conversational style very evident in some sections is deliberate Placing all references in a single bibliography at the end of the text further improves readability. The reader will go to the book to discover a specific fact but be persuaded to read more and derive pleasure from the process. The authors' enthusiasm for the subject comes over strongly in the text, and this enthusiasm proves infectious.
Key Features
* Complement regulators--structure, functional roles and mode of action
* Comprehensive reviews of each of the individual regulators
* Roles of Complement regulators "in vivo, "in health and disease:
* Consequences of deficiency
* Roles in the reproductive system
* Interactions with pathogens
* Exploitation for therapy
* Complement regulators in other species

CRISPR in Animals and Animal Models, Volume 152, the latest release in the Progress in Molecular Biology and Translational Science series, explores the genome editing CRISPR system in cells and animal models, its applications, the uses of the CRISPR system, and the past, present and future of CRISPR genome editing. Topics of interest in this updated volume include a section on CRISPR history, The genome editing revolution, Programming CRISPR and its applications, CRISPR Delivery methods, CRISPR libraries and screening, CRISPR investigation in haploid cells, CRISPR in the generation of transgenic animals, CRISPR therapeutics, and Promising strategies and present challenges.

Natural Variances and Clocks, Volume 99 in the Advances in Genetics series provides the latest information on the rapidly evolving field of genetics, presenting new medical breakthroughs and advances. This updated release includes chapters on a variety of new research, including the Natural variation of the circadian clock in Neurospora, Natural variation and genetics of the photoperiodic timer in the pitcher-plant mosquito, Natural variation in human clocks, and Natural variation in the circadian clock genes in Drosophila and other insects. This series continually publishes important reviews that are ideal for geneticists and their colleagues in affiliated disciplines, critically analyzing future directions.

Peptidomics of Cancer-Derived Enzyme Products, Volume 42, the latest in The Enzymes series, is ideal for researchers in biochemistry, molecular and cell biology, pharmacology, and cancer, with this volume featuring high-caliber, thematic articles on the topic of peptidomics of cancer-derived enzyme products. Specific chapters cover Circulating peptidome and tumor-resident proteolysis, Colon tumor secretopeptidome, Chemoenzymatic method for glycomics, Human plasma peptidome for pancreatic cancer, Lipoproteomics and quantitative proteomics, Salivaomics: Protein markers/extracellular RNA/DNA in saliva, and Enzyme-responsive vectors for cancer therapy.

The dark side of the genome represents vast domains of the genome that are not encoding for proteins - the basic bricks of cellular structure and metabolism. Up to 98% of the human genome is non-coding and produces so-called long non-coding RNA. Some of these non-coding RNA play fundamental roles in cellular identity, cell development and cancer progression. They are now widely studied in many organisms to understand their function. This book reviews this expanding field of research and present the broad functional diversities of those molecules and their putative fundamental and therapeutic roles and develops the recent history of non-coding RNA, their very much debated classification and how they raise a formidable interest for developmental and tumorigenesis biology. Using classical examples and an extensive bibliography, the book illustrates the most studied and attractive examples of these long non-coding RNA, how they interface with epigenetics, genome integrity and expression and what are the current models of their regulatory mechanisms.

The critically acclaimed laboratory standard for more than forty years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. More than 285 volumes have been published (all of them still in print) and much of the material is relevant even today-truly an essential publication for researchersin all fields of life sciences.
Key Features
* Prokaryotic ABC Transporters
* Eukaryotic ABC Transporters
* Nonmammalian ABC Transport Systems
* Mammalian P-Glycoproteins
* Multidrug Resistance Associated Protein
* Cystic Fibrosis Transmembrane Conductance Regulator
* Sulfonylurea Receptor
* Intracellular ABC Transporters

Mechanisms of DNA Recombination and Genome Rearrangements: Methods to Study Homologous Recombination, Volume 600, the latest release in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. Homologous genetic recombination remains the most enigmatic process in DNA metabolism. The molecular machines of recombination preserve the integrity of the genetic material in all organisms and generate genetic diversity in evolution. The same molecular machines that support genetic integrity by orchestrating accurate repair of the most deleterious DNA lesions, however, also promote survival of cancerous cells and emergence of radiation and chemotherapy resistance. This two-volume set offers a comprehensive set of cutting edge methods to study various aspects of homologous recombination and cellular processes that utilize the enzymatic machinery of recombination The chapters are written by the leading researches and cover a broad range of topics from the basic molecular mechanisms of recombinational proteins and enzymes to emerging cellular techniques and drug discovery efforts.

Methods in Enzymology, Volume 599 is the second of two volumes focused on Fe-S cluster enzymes. Topics of interest in this new release include steps towards understanding mitochondrial Fe/S cluster biogenesis, iron sulfur clusters in zinc finger proteins, electrochemistry of Iron-sulfur enzymes, NRVS for Fe in biology and its experiment and basic interpretation, methods for studying iron regulatory protein 1, an important protein in human iron metabolism, the characterization of glutaredoxin Fe-S cluster binding interactions using circular dichroism spectroscopy, fluorescent reporters to track Fe-S cluster assembly and transfer reactions, methods for studying the Fe-S cluster containing base excision repair glycosylase MUTYH, and more.

Volume 2 has focused on aspects of the pituitary gland both anterior (growth hormone and prolactin receptors, and GH action) and posterior (vasopressin) pituitary. In addition, thyroid cancer and steroidogenic enzymes and precocious puberty are covered. Finally, the "hot topics" include leptin and growth factor signaling.

The Many Faces of RNA is the subject for the eighth SmithKline Beecham Pharmaceuticals Research Symposia. It highlights a rapidly developing area of scientific investigation. The style and format are deliberately designed to promote in-depth presentations and discussions and to facilitate the forging of collaborations between academic and industrial partners.
This symposium focuses on several of the many fundamental, advancing strategies for exploring RNA and its functions. It emphasizes the interplay between biology, chemistry, genomics, and molecular biology which is leading to exciting new insights and avenues of investigation. The book explores RNA as a therapeutic target, RNA as a tool, RNA and its interactions, along with chemical, computational, and structural investigations.

Chemical Glycobiology, Part B, Volume 598, the latest release in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume is the second release on chemical glycobiology.

Both eukaryotic and prokaryotic cells depend strongly on the function of ion pumps present in their membranes. The term ion pump, synonymous with active ion-transport system, refers to a membrane-associated protein that translocates ions uphill against an electrochemical potential gradient. Primary ion pumps utilize energy derived from chemical reactions or from the absorption of light, while secondary ion pumps derive the energy for uphill movement of one ionic species from the downhill movement of another species.
In the present volume, various aspects of ion pump structure, mechanism, and regulation are treated using mostly the ion-transporting ATPases as examples. One chapter has been devoted to a secondary ion pump, the Na+-Ca2+ exchanger, not only because of the vital role played by this transport system in regulation of cardiac contractility, but also because it exemplifies the interesting mechanistic and structural similarities between primary and secondary pumps.

A Structure-Function Toolbox for Membrane Transporter and Channels, Volume 594, a new release in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. New chapters in the updated serial include Cryo-EM on membrane proteins embedded in nanodics, Solid-Supported membrane-based electrophysiology on membrane transporters and channels, Saposin-lipoprotein scaffolds for structure determination of membrane transporters and channels, Single-molecule FRET on transporters, Dynamics of channels and transporters investigated by NMR, Structure-function studies on channels and transporters, and a section on MemStar, a new GFP-based expression and purification system for transporters and channels.

Overflow Metabolism: From Yeast to Marathon Runners provides an overview of overflow metabolism, reviewing the major phenomenological aspects as observed in different organisms, followed by a critical analysis of proposed theories to explain overflow metabolism. In our ideal view of metabolism, we think of catabolism and anabolism. In catabolism nutrients break down to carbon dioxide and water to generate biochemical energy. In anabolism nutrients break down to generate building blocks for cell biosynthesis. Yet, when cells are pushed to high metabolic rates they exhibit incomplete catabolism of nutrients, with a lower energy yield and excretion of metabolic byproducts. This phenomenon, characterized by the excretion of metabolic byproducts that could otherwise be used for catabolism or anabolism, is generally known as overflow metabolism. Overflow metabolism is a ubiquitous phenotype that has been conserved during evolution. Examples are the acetate switch in the bacterium E. coli, Crabtree effect in unicellular eukaryote yeasts, the lactate switch in sports medicine, and the Warburg effect in cancer. Several theories have been proposed to explain this seemingly wasteful phenotype. Yet, there is no consensus about what determines overflow metabolism and whether it offers any selective advantage.

G Protein Coupled Receptors, Second Edition, Volume 143, a new volume in the Methods in Cell Biology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. It contains a wide array of topics about the G protein coupled receptors, as well as updates of chapters from the first edition.

This informative publication brings together knowledge of various aspects of cellular regulation. Current Topics in Cellular Regulation reviews the progress being made in those specialized areas of study that have undergone substantial development. It also publishes provocative new theories and concepts and serves as a forum for the discussion of general principles. Researchers in cellular regulation as well as biochemists, molecular and cell biologists, microbiologists, and biophysicists will find Current Topics in Cellular Regulation a useful source of up-to-date information.
Key Features
* Regulation of Iron Metabolism in Eukaryotes
* Regulation of Fas-Mediated Apoptosis
* Aging and Regulation of Apoptosis
* Regulation of Bacterial Responses of Oxidaditive Stress
* Regulation of NF-(B and Disease Control
* Mechanism and Regulation of Bone Resorption
* Gene Regulation by Reactive Oxygen Species
* Structure, Mechanism, and Specificity of Protein-Tyrosine Phosphatases

A proper understanding of the diversity, systematics, and nomenclature of microbes is increasingly important in many branches of biological science. The molecular approach to phylogenetic analysis - pioneered by Carl Woese in the 1970s and leading to the three-domain model (Archaea, Bacteria, Eucarya) - has revolutionized our thinking about evolution in the microbial world. The technological innovation of modern molecular biology and the rapid advancement in computational science have led to a flood of nucleic acid sequence information, bioinformatic tools, and phylogenetic inference methods. Phylogenetic analysis has long played a central role in microbiology and the emerging fields of comparative genomics and phylogenomics require substantial knowledge and understanding of phylogenetic analysis and computational methods. In this book, leading scientists from around the world explore current concepts in molecular phylogeny and their application with respect to microorganisms. The authors describe the different approaches applied today to elucidate the molecular phylogeny of prokaryotes (and eukaryotic protists) and review current phylogenetic methods, techniques, and software tools. Topics covered include: a historical overview, computational tools, multilocus sequence analysis, 16S rRNA phylogenetic trees, rooting of the universal tree of life, applications of conserved indels, lateral gene transfer, endosymbiosis, and the evolution of plastids.

DNA Repair Enzymes, Part A, Volume 591 is the latest volume in the Methods in Enzymology series and the first part of a thematic that focuses on DNA repair enzymes. Topics in this new release include chapters on the Optimization of Native and Formaldehyde iPOND Techniques for Use in Suspension Cells, the Proteomic Analyses of the Eukaryotic Replication Machinery, DNA Fiber Analysis: Mind the Gap!, Comet-FISH for Ultrasensitive Strand-Specific Detection of DNA Damage in Single Cells, Examining DNA Double-Strand Break Repair in a Cell Cycle-Dependent Manner, Base Excision Repair Variants in Cancer, and Fluorescence-Based Reporters for Detection of Mutagenesis in E. coli.