The recent advent of molecular technologies has changed the way plant breeders identify and select their germplasm as genetic variation can now be assessed at the DNA level. Crop Breeding: Methods and Protocols presents detailed guidelines and tutorials that suit different needs and capacity from small laboratories analyzing molecular markers on a one-by-one basis to the increasingly popular high-throughput protocols for high capacity laboratories. Topics covered include breeding strategy for the selection of an ideal variety or genetic ideo type, protocols for breeders using molecular markers in selection programs and for laboratories providing molecular services to breeding programs, statistical programs and software to aid implementation of molecular data into breeding programs and methodologies that facilitate the generation of genetic diversity and its characterization. Written in the 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 protocols and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Crop Breeding: Methods and Protocols will help in expanding the use of molecular technologies for the creation of tomorrow's crop varieties.

Every cell in our bodies contains a great variety and number of permeability pathways for various organic and inorganic ions, water, metabolites, nutrients, and signalling molecules. Maintenance and precise control of gating within these pathways are fundamental principles of life as these underlie basic cellular functions such as communication, contractility, and metabolism. In Ion Channels: Methods and Protocols, Second Edition, experts in the field contribute chapters that focus on the strategies, approaches, methods, and protocols for studying a large family of proteins that form ionic channels in the plasma membrane and intracellular membranes of cells. Using practical examples from the cutting-edge current research, this volume will take a look back at the major methods and approaches that aided the progress toward the current understanding of ion channel function, structural design, and biological roles. The volume also aims to look forward and identify approaches that will lead us to future discoveries. Written in the highly successful Methods in Molecular Biology (TM) 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, Ion Channels: Methods and Protocols, Second Edition will greatly assist researchers searching for specific methodology in studying ion channels on the path toward a greater understanding of these key biological features.

Systems biology can now be considered an established and fundamental field in life sciences. It has moved from the identification of molecular 'parts lists' for living organisms towards synthesising information from different 'omics'-based approaches to generate and test new hypotheses about how biological systems work. In In Silico Systems Biology: Methods and Protocols, expert researchers in the field detail a practical set of chapters based often on actual materials used and develop for face-to-face training with examples and case studies. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, step-by-step workflows, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, In Silico Systems Biology: Methods and Protocols seeks to aid scientists in the further study of network biology and mathematical models of biological systems.

The Endoplasmic Reticulum (ER) is an organelle with extraordinary signaling and homeostatic functions. It is the organelle responsible for protein folding, maturation, quality control and trafficking of proteins destined for the plasma membrane or for secretion into the extracellular environment. Failure, overloading or malfunctioning of any of the signaling or quality control mechanisms occurring in the ER may provoke a stress condition known as ER stress . Accumulating evidence indicates that ER stress may dramatically perturb interactions between the cell and its environment, and contribute to the development of human diseases, ranging from metabolic diseases and cancer to neurodegenerative diseases, or impact therapeutic outcome. This book primarily focuses on the pathophysiology of ER stress. It introduces the molecular bases of ER stress, the emerging relevance of the ER-mitochondria cross-talk, the signaling pathways engaged and cellular responses to ER stress, including the adaptive Unfolded Protein Response (UPR), autophagy as well as cell death. Next the book addresses the role of ER stress in physiology and in the etiology of relevant pathological conditions, like carcinogenesis and inflammation, neurodegeneration and metabolic disease. The last chapter describes how ER stress pathways can be targeted for therapeutic benefit. Altogether, this book will provide the reader with an exhaustive view of ER stress biology and the latest insights in the role of ER stress in relevant human diseases."

How do you keep track of basic information on the proteins you work with? Where do you find details of their physicochemical properties, amino acid sequences, and structure? Are you tired of scanning review articles, primary papers, and databases to locate that elusive fact?
The Academic Press FactsBook Series has established itself as the best source of easily-accessible and accurate facts about protein groups. Described as "a growing series of excellent manuals" by "Molecular Medicine Today" and "essential works of reference" by "Trends in Biochemical Sciences," the FactsBooks have become the most popular comprehensive data resources available. Using an easy-to-follow format, the FactsBooks will keep you up-to-date with the latest advances in structure, amino acid sequences, physicochemical properties, and biological activity. Meticulously researched and compiled by experts in the field, keeping abreast of developments has never been so easy
The Transporter FactsBook contains entries covering almost 800 transporters. Organized into 55 families of structurally related transporters, this volume includes ATPases, ABC transporters, H+-dependent antiporters and symporters, Na+-dependent antiporters and symporters, and other transporters such as mitochondrial transporters.
Key Features
* Nomenclature, biological sources and substrates
* Phylogenetic relationships
* Predicted structures
* Physical and genetic characteristics
* Multiple amino acid sequence alignments
* Database accession numbers
* Key references

Researchers in the field of ecological genomics aim to determine how a genome or a population of genomes interacts with its environment across ecological and evolutionary timescales. Ecological genomics is trans-disciplinary by nature. Ecologists have turned to genomics to be able to elucidate the mechanistic bases of the biodiversity their research tries to understand. Genomicists have turned to ecology in order to better explain the functional cellular and molecular variation they observed in their model organisms.

We provide an advanced-level book that covers this recent research and proposes future development for this field. A synthesis of the field of ecological genomics emerges from this volume."Ecological Genomics"covers a wide array of organisms (microbes, plants and animals) in order to be able to identify central concepts that motivate and derive from recent investigations in different branches of the tree of life.

"Ecological Genomics"covers 3 fields of research that have most benefited from the recent technological and conceptual developments in the field of ecological genomics: the study of life-history evolution and its impact of genome architectures; the study of the genomic bases of phenotypic plasticity and the study of the genomic bases of adaptation and speciation.

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Gaining an understanding of the mechanisms by which cells process and respond to extracellular cues has become a major goal in many areas of bi- ogy and has attracted the attentions of almost every traditional discipline within the biological sciences. At the heart of these divergent endeavors are common methods that can aid biochemists, physiologists, and pharmacologists in ta- ling the specific questions addressed by their research. In Receptor Signal Transduction Protocols, a diverse array of meth- ologies employed to interrogate ligand--receptor and receptor-effector int- actions are described by authors who have devised and successfully applied them. The authors blend excellent descriptions and applications of fairly well established methodologies with new technologies at the cutting-edge of signal transduction research and as such I hope the present volume will complement and extend a previous excellent volume in this series edited by David Kendall and Stephen Hill (Methods Molecular Biology, vol. 41, Signal Transd- tion Protocols).

Protein informatics is a newer name for an already existing discipline. It encompasses the techniques used in bioinformatics and molecular modeling that are related to proteins. While bioinformatics is mainly concerned with the collection, organization, and analysis of biological data, molecular modeling is devoted to representation and manipulation of the structure of proteins.

Protein informatics requires substantial prerequisites on computer science, mathematics, and molecular biology. The approach chosen here, allows a direct and rapid grasp on the subject starting from basic knowledge of algorithm design, calculus, linear algebra, and probability theory.

An Introduction to Protein Informatics, a professional monograph will provide the reader a comprehensive introduction to the field of protein informatics. The text emphasizes mathematical and computational methods to tackle the central problems of alignment, phylogenetic reconstruction, and prediction and sampling of protein structure.

An Introduction to Protein Informatics is designed for a professional audience, composed of researchers and practitioners within bioinformatics, molecular modeling, algorithm design, optimization, and pattern recognition. This book is also suitable as a graduate-level text for students in computer science, mathematics, and biomedicine.

Radiation Oncology and Radiotherapy, Part A, Volume 172 in the Methods in Cell Biology series, highlights advances in the field, with this new volume presenting interesting chapters on timely topics, including DNA damage quantification by the COMET assay, Immunofluorescence microscopy-assisted quantification of ATM and ATR activation in irradiated cells, Immunoblotting-based characterization of the DNA damage response, Assessment of lipid peroxidation in irradiated cells, A simple method to assess clonogenic survival of irradiated cancer cells, Quantification of beta-galactosidase activity as a marker of radiation-driven cellular senescence, Cytofluorometric assessment of cell cycle progression in irradiated cells, and more. Other sections cover Assessment of transcription inhibition as a characteristic of immunogenic cell death, Assessment of eIF2a phosphorylation during immunogenic cell death, Quantification of cytosolic DNA species by immunofluorescence and automated image analysis, Flow cytometry-assisted quantification of CALR exposure during immunogenic cell death, Interference of immunogenic anticancer therapy by artificially controlled calreticulin secretion from tumor cells, along with many additional topics of interest.

Over the last decade the development of new molecular biology tools, advanced microscopy, live imaging and systems biology approaches have revolutionized our conception of how embryonic development proceeds. One fundamental aspect of development biology is the concept of morphogenesis: understanding how a group of multipotent cells organize and differentiate into a complex organ. In Kidney Development: Methods and Protocols, expert researchers in the field detail different approaches to tackle kidney development. These approaches include culture and live imaging aspects of kidney development, analyzing the 3-dimensional aspects of branching morphogenesis as well as nephrogenesis, manipulation of the gene/protein expression during kidney development as well as in the adult kidney, and how to assess kidney malformation and disease. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Kidney Development: Methods and Protocols seeks to aid scientists in the further study of the process of morphogenesis which is fundamental important not only for studying developmental biology but also for regenerative medicine.

During the course of evolution, an imbalance was created between the rate of vertebrate genetic adaptation and that of the lower forms of living organisms, such as bacteria and viruses. This imbalance has given the latter the advantage of generating, relatively quickly, molecules with unexpected structures and features that carry a threat to vertebrates. To compensate for their weakness, vertebrates have accelerated their own evolutionary processes, not at the level of whole organism, but in specialized cells containing the genes that code for antibody molecules or for T-cell receptors. That is, when an immediate requirement for molecules capable of specific interactions arose, nature has preferred to speed up the mode of Darwinian evolution in pref- ence to any other approach (such as the use of X-ray diffraction studies and computergraphic analysis). Recently, Darwinian rules have been adapted for test tube research, and the concept of selecting molecules having particular characteristics from r- dom pools has been realized in the form of various chemical and biological combinatorial libraries. While working with these libraries, we noticed the interesting fact that when combinatorial libraries of oligopeptides were allowed to interact with different selector proteins, only the actual binding sites of these proteins showed binding properties, whereas the rest of the p- tein surface seemed "inert. " This seemingly common feature of protein- having no extra potential binding sites--was probably selected during evolution in order to minimize nonspecific interactions with the surrounding milieu.

The endoplasmic reticulum (ER) is an extensive network of membranes that folds, modifies, and transports proteins in eukaryotic cells. It also manufactures lipids and interacts extensively with other organelles, playing essential roles in cell growth and homeostasis. Written and edited by experts in the field, this collection from Cold Spring Harbor Perspectives in Biology covers all aspects of ER morphology and function, as well as its interactions with the nucleus, Golgi, and mitochondria. Contributors examine how proteins translocate across the ER membrane, the processes that occur inside the ER lumen (e.g., folding, glycosylation, and disulfide bond formation), and how the proteins are packaged into vesicles and transported to the Golgi. They also review quality-control mechanisms that are employed by the ER to detect and eliminate misfolded or unassembled proteins. Lipid synthesis and transport are also discussed. This volume covers not only the biochemistry and cell biology of the ER, but also ER stress, metabolism, and the role of the ER in viral replication. Thus, it is an essential reference for cell biologists, physiologists, and pathologists interested in understanding the numerous functions of the ER.

From basic performing of sequence alignment through a proficiency at understanding how most industry-standard alignment algorithms achieve their results, "Multiple Sequence Alignment Methods" describes numerous algorithms and their nuances in chapters written by the experts who developed these algorithms. The various multiple sequence alignment algorithms presented in this handbook give a flavor of the broad range of choices available for multiple sequence alignment generation, and their diversity is a clear reflection of the complexity of the multiple sequence alignment problem and the amount of information that can be obtained from multiple sequence alignments. Each of these chapters not only describes the algorithm it covers but also presents instructions and tips on using their implementation, as is fitting with its inclusion in the highly successful "Methods in Molecular Biology" series.

Authoritative and practical, "Multiple Sequence Alignment Methods" provides a readily available resource which will allow practitioners to experiment with different algorithms and find the particular algorithm that is of most use in their application."

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.

The last decade has witnessed the unravelling of remarkable new insights into the biology of Vibrio cholerae. These include the discovery of the filamentous phage that encodes cholera toxin, the existence of two chromosomes in V. cholerae, and the sequencing of the whole genome of the V. cholerae O1 strain N16961. These pioneering works have led to an inevitable escalation in the amount of data generated. This book distills the essence of this mighty deluge of information, providing a timely review of the genomics and molecular biology of this important human pathogen. Written by leading V. cholerae experts, the chapters review the most important cutting-edge genetic facets of V. cholerae including its genomic organization, population genetics, molecular epidemiology, and synchronized regulation of gene expression. Other topics include the molecular basis for enhanced transmissibility of cholera during epidemics, survival of the pathogen in the environment, and above all the evolution

Molecular modeling has undergone a remarkable transformation in the last 20 years. This book provides thorough introductions and a compilation of step-by-step methods applicable to problems faced by non-specialists especially those new to the software packages used in molecular modeling. Tips on troubleshooting and avoiding common pitfalls are included in the book, along with chapters covering a wide range of subjects. Links to downloadable software are also provided.

Neuronal Nicotinic Receptors: Pharmacology and Therapeutic Opportunities
Edited by Stephen P. Arneric and Jorge D. Brioni
Nicotine's efficacy in the treatment of pain and anxiolysis, and its usefulness as a memory enhancer, have been suspected for many years, but research into the molecular biology of nicotinic acetylcholine receptors (nAChRs) traditionally has focused on smoking cessation. Now, thanks to recent technological advances in the molecular biology and electrophysiology of nAChRs, an abundance of evidence has emerged in support of nAChR ligands in the treatment of a range of CNS disorders, including Alzheimer's disease, Parkinson's disease, neuropathic pain, attention deficit disorder, and depression.
Featuring contributions from an international team of experts in the field, this volume provides researchers and clinicians with a comprehensive review of nAChRs, their molecular biology, pharmacology, and clinical implications. Moving from the molecular to the clinical levels, the authors explore the latest developments in all areas of nAChR research, including:
* Molecular biology and biochemistry of nAChRs
* The physiological roles of nAChRs
* Pharmacokinetics of nicotine-like alkaloids
* Recent breakthroughs and emerging trends in nAChR medicinal chemistry
* Current and emerging therapeutic applications of novel nAChR ligands.

A timely, authoritative review of basic research into nAChRs and their role in the development of new therapeutic strategies in the treatment of a range of various CNS diseases, Neuronal Nicotinic Receptors: Pharmacology and Therapeutic Opportunities is required reading for neuroscientists, pharmacologists, medicinal chemists, biological psychiatrists, and psychopharmacologists.

A large number of newly-synthesized polypeptides must cross one or several intracellular membranes to reach their functional locations in the eukaryotic cell. The mechanisms of protein trafficking, in particular the post-translational targeting and membrane translocation of proteins, are of fundamental biological importance and are the focus of intensive research world-wide. For more than 15 years, mitochondria have served as the paradigm organelle system to study these processes. Although key questions, such as how precisely proteins cross a membrane, still remain to be answered, exciting progress has been made in understanding the basic pathways of protein import into mitochondria and the components involved. In addition to a fascinating richness and complexity in detail, the analysis of mitochondrial protein import has revealed mechanistic principles of general significance: Major discoveries include the demonstration of the requirement of an unfolded state for translocation and of the essential role of molecular chaperones on both sides of the membranes in maintaining a translocation-competent conformation and in protein folding after import. It is becoming clear how a polypeptide chain is "reeled" across the membrane in an ATP-dependent process by the functional cooperation of membrane proteins, presumably constituting part of a transmembrane channel, with peripheral components at the trans-side of the membrane.
In this volume, eminent experts in the field take the time to review the central aspects of mitochondrial biogenesis. The logical order of the 16 chapters is determined by the sequence of steps during protein import, starting with the events taking place in the cytosol, followed by the recognition of targeting signals, the translocation of precursor proteins across the outer and inner membranes, their proteolytic processing and intramitochondrial sorting, and finally their folding and oligomeric assembly. In addition, the mechanisms involved in the export of mitochondrially encoded proteins as well as recent advances in understanding the division and inheritance of mitochondria will be discussed.

The field of DNA repair is vast and advancing rapidly. Recent investigations have begun to focus on the involvement of chromatin in the repair of broken DNA. Although I have no doubt that many breakthroughs in our understanding of chromatin, chromatin regulation, and DNA repair lie in our future, presently this is a new line in inquiry. As such there are many, many unanswered questions. Indeed, most of the correct questions have probably not even been asked yet. Here I have attempted to present a review of some of the current body of knowledge that may prove relevant to understanding the role of chromatin in DNA repair. Because the volume of research, and the relevant findings, come from a staggering array of labs, systems, and ideas I have focused primarily on findings developed from the study of the budding yeast Saccharomyces cerevisiae. Unfortunately, this means that I have left out a great deal of information. It is my hope, however, that the information I do detail, particularly in Chapter 1, will give a flavor for the scope of the problem and perhaps highlight some of the interesting directions this field is taking, or may one day take. I would also point out that the primary research that is presented herein is not in any way meant to represent the comprehensive scope of research being performed. To understand DNA repair will require investigation from innumerable labs, performed by innumerable researchers, moving in unexpected directions.

It is now clear from a wide range of research that cytoplasm is not merely a buffered solution of proteins and enzymes but contains a series of complex filamentous structures. The cytoskeleton is the collective term given to these filaments. There is a considerable amount of data available on the protein composition of the major filament systems (microfilaments, microtubules, and intermediate filaments) but we are still comparatively ignorant about the role of the cytoskeleton in cell physiology. However such major cytoplasmic components (actin and tubulin, the monomeric constituents of microfilaments and microtubules, are major cell proteins) must have important roles to play in cell function, and investigations into the functional role of the cytoskeleton currently represent a major area of cell biological research.
In recent years rapid advances in molecular biology have begun to influence research on the cytoskeleton. This trend is sure to continue and the techniques of molecular biology and genetics are set to make major contributions to our understanding of the cytoskeleton, as illustrated in this volume by several reviews; the use of transfection techniques by Ben-Ze'ev, the power of Drosophila genetics is described by Fyrberg and the major advances made in the inesin field using molecular approaches as described by Cyr et al. The chapters by Fyrberg and Cyr et al. also illustrate two other areas where major advances in our understanding of the cytoskeleton is occuring; the great array of different motor proteins involved in intracellular movements and the study of the cytoskeleton in developmental biology.
Overall, Volume 12 in this series illuminates our increasing knowledge of the important roles of the cytoskeleton in cell function, particularly how it is central to metabolic organization, intracellular transport, interactions with matrix, and nerve function. Our knowledge of the cytoskeleton is now reaching a stage where it is clear that abnormalities in the organization of the cytoskeleton can lead to important clinical manifestations of disease; an example of how such research is now impinging on medical science is presented in the final chapter by Lane on keratin diseases.

Written by specialists in the field, this book provides an overview of the structural and molecular biology of cellular processes that occur at or near bacterial membranes.

Drawing especially on insights emerging from studies of the cellular networks formed by fungi, this book describes the fundamental indeterminacy that enables life forms to thrive in and create inconstant circumstances. It explains how indeterminacy arises from counteraction between associative and dissociative processes at the reactive interfaces between living systems and their surroundings. It stresses the relevance of these processes to understanding the dynamic contexts within which living systems of all kinds - including human societies-explore for, use up, conserve and recycle sources of energy.By focusing on dynamic boundaries, the book counterbalances the discretist view that living systems are assembled entirely from building-block-like units - individuals and genes - that can be freely sifted, as opposed to channeled, by natural selection. It also shows how the versatility that enables life forms to proliferate in rich environments, whilst minimizing losses in restrictive environments, depends on capacities for error and co-operation within a fluid, non-hierarchical power structure. Understanding this point yields a more compassionate, less competitive and less self-centred outlook on life's successes and failures.

Posttranslational Modifications of Proteins: Tools for Functional Proteomics is a compilation of detailed protocols needed to detect and analyze the most important co- and posttranslational modifications of proteins. Though, for reasons of simplicity not explicitly mentioned in the title, both kinds of modifications are covered, whether they occur during, or after, biosynthesis of the protein. My intention was to cover the most significant protein modifications, focusing on the fields of protein function, proteome research, and the characterization of pharmaceutical proteins. The majority of all proteins undergo co- and/or posttranslational modifications. Knowledge of these modifications is extremely important, since they may alter physical and chemical properties, folding, conformation distribution, stability, act- ity, and, consequently, function of the proteins. Moreover, the modification itself can act as an added functional group. Examples of the biological effects of protein mo- fications include: phosphorylation for signal transduction, ubiquitination for p- teolysis, attachment of fatty acids for membrane anchoring or association, glycosylation for protein half-life, targeting, cell-cell and cell-matrix interactions, and carboxylation in protein-ligand binding to name just a few. Full understanding of a specific protein structure-function relationship requires detailed information not only on its amino acid sequence, which is determined by the corresponding DNA sequence, but also on the presence and structure of protein modifications.

This important volume is mainly concerned with the development of methods for "sequencing" - that is, determination of the order of the amino acids in proteins and of nucleotides in RNA and DNA. In 1943 the position of only one amino acid in a protein (insulin) was known, and Sanger's first paper resulted in finding a second amino acid. In his final paper in 1982 he describes the determination of a DNA sequence of 48,502 nucleotides. The papers describe the steady improvements in techniques, and exciting biological results revealed by the sequences.