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.

This book is authored by an exciting mixture of top experts and young rising stars from the fields of molecular chaperones and stress adaptation. In addition to giving a comprehensive summary with original references to their field, all authors share their hypotheses and vision on future trends with the reader. The book makes a novel synthesis of the molecular aspects of the stress response and long term adaptation processes with the system biology approach of biological networks. A novel perspective of old facts is provided in each chapter, where old means only 5-10 years in this rapidly expanding field. The integrative, holistic nature of the book makes it an excellent reading for undergraduates, graduate students and fellow scientists extending their knowledge and studies.

Do real stem cells and stem cell lineages exist in lower organisms? Can stem cells from one organism parasitize the soma and/or the germ line of conspecifics? Can differentiated cells in marine organisms be re-programmed to regenerate tissues, organs and appendages through novel de-differentiation, transdifferentiation, or re-differentiation processes, leading to virtually all three germ layers, including the germline? The positive answers to above questions open a new avenue in stem cell research: the biology of stem cells in marine organisms. It is therefore unfortunate that while the literature on stem cell from terrestrial organisms is rich and expanding at an exponential rate, investigations on marine organisms stem cells are very limited and scarce.

By presenting theoretical chapters, overview essays and specific research results, this book summarises the knowledge and the hypotheses on stem cells in marine organisms through major phyla and specific model organisms. The study on stem cells from marine invertebrates may shed lights on mechanisms promoting immunity, developmental biology, regeneration and budding processes in marine invertebrates, body maintenance, aging and senescence. It aims in encouraging a larger scientific community to follow and study the novel phenomena of stem cells behaviours as depicted from the few currently studied marine invertebrates.

QTL Mapping in Crop Improvement: Present Progress and Future Perspectives presents advancements in QTL breeding for biotic and abiotic stresses and nutritional improvement in a range of crop plants. The book presents a roadmap for future breeding for resilience to various stresses and improvement in nutritional quality. Crops such as rice, wheat, maize, soybeans, common bean, and pigeon pea are the major staple crops consumed globally, hence fulfilling the nutritional requirements of global populations, particularly in the under-developed world, is extremely important. Sections cover the challenges facing maximized production of these crops, including diseases, insect damage, drought, heat, salinity and mineral toxicity. Covering globally important crops including maize, wheat, rice, barley, soybean, common bean and pigeon pea, this book will be an important reference for those working in agriculture and crop improvement.

The fungi have been major players in the molecular revolution that has transformed biology. Because they can be manipulated as microorganisms, yeast and Neurospora provide information that is difficult to acquire with plants and animals, and experimental findings with fungi often throw light on corresponding processes in plants and animals. The filamentous fungus Neurospora crassa has become a valuable model organism because of its favorable features for genetic analysis and because of the vast store of information that has been acquired during 75 years of research. This compendium provides researchers and students with a concise account of current knowledge about the genes and genome of Neurospora, setting the stage for research that will follow completion of the genome sequence.
This book, which is fully documented and abundantly illustrated, will be an indispensable tool in any laboratory that uses fungi for research in molecular genetics, classical genetics, developmental genetics, or cell biology.
Key Features
* Molecular, genetic, and phenotypic information for over 1000 nuclear genes
* Genetic maps.
* Linkage group assignments for 1000 loci
* 2300 references, 68 figures
* Guide to electronic and other sources of information
* Summary information on the mitochondrial genome
* cDNAs identified from different stages of life
* Classical, cytogenetic, and molecular data, anticipating completion of the genome sequence

From New York Times bestselling author Sam Kean comes more incredible stories of science, history, language, and music, as told by our own DNA.
In "The Disappearing Spoon," bestselling author Sam Kean unlocked the mysteries of the periodic table. In THE VIOLINIST'S THUMB, he explores the wonders of the magical building block of life: DNA.
There are genes to explain crazy cat ladies, why other people have no fingerprints, and why some people survive nuclear bombs. Genes illuminate everything from JFK's bronze skin (it wasn't a tan) to Einstein's genius. They prove that Neanderthals and humans bred thousands of years more recently than any of us would feel comfortable thinking. They can even allow some people, because of the exceptional flexibility of their thumbs and fingers, to become truly singular violinists.
Kean's vibrant storytelling once again makes science entertaining, explaining human history and whimsy while showing how DNA will influence our species' future.

This volume contains state-of-the-art methods tackling all aspects of small non-coding RNAs biology. Small Non-Coding RNAs: Methods and Protocols guides readers through customized dedicated protocols and technologies that will be of valuable help to all those willing to contribute deciphering the numerous functions of small non-coding RNAs. Written in the highly successful Methods of 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 key tips on troubles troubleshooting and avoiding known pitfalls. Instructive and practical, Small Non-Coding RNAs: Methods and Protocols reaches out to biochemists, cellular and molecular biologists already working in the field of RNA biology and to those just starting to study small non-coding RNAs.

Completion of the sequence of the human genome represents an unpar- leled achievement in the history of biology. The project has produced nearly complete, highly accurate, and comprehensive sequences of genomes of s- eral organisms including human, mouse, drosophila, and yeast. Furthermore, the development of high-throughput technologies has led to an explosion of projects to sequence the genomes of additional organisms including rat, chimp, dog, bee, chicken, and the list is expanding. The nearly completed draft of genomic sequences from numerous species has opened a new era of research in biology and in biomedical sciences. In keeping with the interdisciplinary nature of the new scientific era, the chapters in Gene Mapping, Discovery, and Expression: Methods and Protocols recapitulate the necessity of integration of experimental and computational tools for solving - portant research problems. The general underlying theme of this volume is DNA sequence-based technologies. At one level, the book highlights the importance of databases, genome-browsers, and web-based tools for data access and ana- sis. More specifically, sequencing projects routinely deposit their data in p- licly available databases including GenBank, at the National Center of Biotechnology (NCBI) in the United States; EMBL, maintained by the European Bioinformatics Institute; and DDBJ, the DNA Data Bank of Japan. Currently, several browsers offer facile access to numerous genomic DNA sequences for gene mapping and data retrieval.

Oxidation is any reaction in which electrons are removed from a molecule, thus increasing the number of binding sites on the molecule that are able to react with other atoms and molecules.;This volume addresses oxidant-reduction or redox and antioxidant sensitive molecular mechanisms and how they are implicated in different disease processes. Recent work in this area has revealed that these mechanisms may be linked with different disease processes, such as immune response, cell proliferation, inflammation, metabolism, ageing and cell death. Possible strategies to pharmacologically and/or nutritionally manipulate such redox-sensitive molecular responses are emphasized.

This volume describes high-throughput approaches to a series of robust, established methodologies in molecular genetic studies of population samples. Such developments have been essential not only to linkage and association studies of single-gene and complex traits in humans, animals and plants, but also to the characterisation of clone banks, for example in mapping of genomes. Chapters have been written by developers or highly experienced end-users concerned with a diverse array of biological applications. The book should appeal to any researcher for whom costs and throughput in their genetics laboratory have become an issue.

Genomics is a new and fast expanding area of biology encompassing high throughput or large scale experimentation at the whole genome level, and the organization, analysis and interpretation of the huge amount of data emerging from genome projects. Major new technologies have evolved recently that enable experimentation at the whole genome level, and more novel technologies are currently being developed. This volume describes in detail the new technology necessary to study the entire genome in a holistic manner and all the high throughput and large-scale experimental methodologies currently being used in genomic science. In addition the authors describe the progress of the newest technologies that are currently being developed. Written by experts in the field, this concise yet informative volume covers all aspects of technology pertaining to genomic studies. It is an essential book for anyone involved in genomic science.

This revised edition provides up-to-date protocols developed in the HSC field. A team of leading researchers supply this volume with in-depth, readily reproducible methods for effective characterization of HSC and their developmental potential. The book provides detailed flow cytometry protocols for thorough analysis of enriched HSC populations, and offers a variety of transplantation approaches to measure HSC function in vivo. This is a much needed technical resource in the critically important field of stem cell investigation.

Plasmids are fascinating entities which can replicate autonomously in bacterial, archaeal, and eukaryotic cells. They profit from the cellular environment of the host but can also carry a rich diversity of genes which can be beneficial for the host. Plasmids confer the ability to degrade organic compounds and to fix nitrogen. In addition, plasmids carry antibiotic resistance genes and their spread in pathogenic bacteria is of great medical importance. Plasmids are used in molecular studies of various organisms with ramifications in synthetic biology, medicine, ecology, and evolution, as well as basic research in molecular and structural biology. Written by acknowledged experts in the field, this volume provides an up-to-date treatment of the structure, function, and application of plasmids, with a particular emphasis on current and future trends. The book is aimed primarily at research scientists, graduate students, and professional scientists, but will also be of great interest to all

Antisense technology is the ability to manipulate gene expression within mammalian cells providing powerful experimental approaches for the study of gene function and gene regulation. For example, methods that inhibit gene expression permit studies which probe the normal function of a specific product within a cell. Such methodology can be used in many disciplines such as pharmacology, oncology, genetics, cell biology, developmental biology, molecular biology, biochemistry, and neurosciences. This volume will be a truly important tool in biomedical-oriented research.
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. Now with more than 300 volumes (all of them still in print), the series contains much material still relevant today--truly an essential publication for researchers in all fields of life sciences.

Although embryonic stem cells currently enjoy the public limelight and show great pr- ise for cell based medical therapies, it is the adult stem cells which are responsible for the body's natural ability to fght disease, heal and recover, or fail and succumb to various maladies. The study of mammalian adult stem cells has surged recently, most likely from a maturation of stem cell studies in the classical developmental model organisms and in hematopoeisis. All the tissues of the body examined so far are generated and regenerated from stem cells, it has been an important frst step to adapt or devise new methods to identify and obtain these cells in quantity and purity for further study. Culture techniques have been optimized for managing the growth and differentiation of stem cells in vitro; as some stem cells are pluripotent, often the method is to guide the fate of such cells among the possible differentiation fates. Much of this work, and that in the classical model org- isms, has helped defne the aspects of the stem cell environment or niche that are crucial for both growth and differentiation, and these studies have moved in vivo at increasingly higher resolution. Importantly, the in vivo niche is a current target for bioengineering the matrix and signaling factors. Herein, we present methods for studying six types of mammalian stem cells, m- mary, neural, mesenchymal, endothelial, dendritic, and muscle.

Computer access is the only way to retrieve up-to-date sequences and this book shows researchers puzzled by the maze of URLs, sites, and searches how to use internet technology to find and analyze genetic data. The book describes the different types of databases, how to use a specific database to find a sequence that you need, and how to analyze the data to compare it with your own work.
The content also covers sequence phenotype, mutation, and genetic linkage databases; simple repetitive DNA sequences; gene feature identification; and prediction of structure and function of proteins from sequence information. This book will be invaluable to those starting a career in life sciences research as well as to established researchers wishing to make full use of available resources.
Key Features
* Describes a wide range of databases: DNA, RNA, protein, pathways, and gene expression
* Enables readers to access the information they need from databases on the web
* Includes a directory of URLs for easy reference
* Invaluable for those starting a career in life sciences research and also for established researchers wishing to make full use of available resources.

As microarray technology has matured, data analysis methods have advanced as well. Methods Of Microarray Data Analysis III is the third book in this pioneering series dedicated to the existing new field of microarrays. While initial techniques focused on classification exercises (volume I of this series), and later on pattern extraction (volume II of this series), this volume focuses on data quality issues. Problems such as background noise determination, analysis of variance, and errors in data handling are highlighted.

Three tutorial papers are presented to assist with a basic understanding of underlying principles in microarray data analysis, and twelve new papers are highlighted analyzing the same CAMDA'02 datasets: the Project Normal data set or the Affymetrix Latin Square data set. A comparative study of these analytical methodologies brings to light problems, solutions and new ideas. This book is an excellent reference for academic and industrial researchers who want to keep abreast of the state of art of microarray data analysis.

This second edition volume discusses the revolutionary development of faster and less expensive DNA sequencing technologies from the past 10 years and focuses on general technologies that can be utilized by a wide array of plant biologists to address specific questions in their favorite model systems. This book is organized into five parts. Part I examines the tools and methods required for identifying epigenetic and conformational changes at the whole-genome level. Part II presents approaches used to determine key aspects of a gene's function, such as techniques used to identify and characterize gene regulatory networks. This is followed by a discussion of tools used to analyze the levels of mRNA, mRNA translation rates and metabolites. Part III features a compilation of forward and reverse genetic approaches that include recent implementation of high-throughput sequencing in classical methodologies such as QTL mapping. The final two parts explore strategies to facilitate and accelerate the generation and testing of functional DNA elements and basic computational tools used to facilitate the use of systems biology approached by a broad spectrum of plant researchers. Written in the highly successful Methods of 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 key tips on troubleshooting and avoiding known pitfalls. Practical and timely, Plant Functional Genomics: Methods and Protocols, Second Edition highlights the latest developments in DNA sequencing technologies that are likely to continue shaping the future of functional genomics.

The temptations of a new genetically informed eugenics and of a revived faith-based, world-wide political stance, this study of the interaction of science, religion, politics and the culture of celebrity in twentieth-century Europe and America offers a fascinating and important contribution to the history of this movement. The author looks at the career of French-born physician and Nobel Prize winner, Alexis Carrel (1873-1944), as a way of understanding the popularization of eugenics through religious faith, scientific expertise, cultural despair and right-wing politics in the 1930s and 1940s. Carrel was among the most prestigious experimental surgeons of his time who also held deeply illiberal views. In Man, the Unknown (1935), he endorsed fascism and called for the elimination of the unfit. The book became a huge international success, largely thanks to its promotion by Readers' Digest as well as by the author's friendship with Charles Lindbergh. In 1941, he went into the service of the French pro-German regime of Vichy, which appointed him to head an institution of eugenics research. His influence was remarkable, affecting radical Islamic groups as well Le Pen's Front National that celebrated him as the founder of ecology. It includes a foreword by Herman Lebovics.

As researchers continue to make enormous progress in mapping disease genes, exciting, novel, and complex analyses have emerged. In this book, scientists from around the world, who are leaders in this field, contribute their vast experience and expertise to produce a comprehensive and fascinating text for researchers and clinicians alike. They provide cutting-edge analysis of the most up-to-date and preeminent information available.

In this book leading researchers in the field discuss the state-of-the-art of many aspects of SAPK signaling in various systems from yeast to mammals. These include various chapters on regulatory mechanisms as well as the contribution of the SAPK signaling pathways to processes such as gene expression, metabolism, cell cycle regulation, immune responses and tumorigenesis. Written by international experts, the book will appeal to cell biologists and biochemists.

Hepatobiliary cancer refers to primary malignant tumors originating in cells of the liver, bile ducts, and gallbladder. Globally, primary liver cancer, which includes hepatocellular carcinoma (~75 % of all cases) and intrahepatic biliary cancer or cholangiocarcinoma (~10-15 % 0f all cases) is the 6th most commonly diagnosed cancer and 3rd leading cause of cancer deaths worldwide. The vast majority of these highly malignant cancers are diagnosed at an advanced stage where treatment options are limited and patient survival outcomes are poor. The biological and therapeutic challenges posed by hepatobililiary cancers such as hepatocellular carcinoma (HCC) and cholangiocarcinoma (CCA) are daunting, emphasizing a critical need to review and assess current and evolving basic, translational, and clinical research focused on addressing the critical obstacles that continue to limit progress towards achieving significant improvements in HCC and CCA clinical management and patient survival outcomes. Towards this goal, this special edition of Advances in Cancer Research is focused on providing a comprehensive, timely and authoritative reviews covering such topics of significant scientific and clinical relevance, including hepatobiliary cancer risk mechanisms and risk-predictive molecular biomarkers; causes and functional intricacies of inter- and intratumor heterogeneity; novel insights into the role of tumor microenvironment and key signaling pathways in promoting hepatobiliary cancer progression, therapeutic resistance and immunosuppression; emerging biomarkers of HCC and CCA prognosis; advances in molecular genomics for personalizing tumor classification and targeted therapies; innovative preclinical cell culture modeling for hepatobiliary cancer drug discovery; and current and emerging trends in hepatobiliary cancer molecular therapeutic targeting and immunotherapies.

Most organisms and populations have to cope with hostile environments, threatening their existence. Their ability to respond phenotypically and genetically to these challenges and to evolve adaptive mechanisms is, therefore, crucial. The contributions to this book aim at understanding, from a evolutionary perspective, the impact of stress on biological systems. Scientists, applying different approaches spanning from the molecular and the protein level to individuals, populations and ecosystems, explore how organisms adapt to extreme environments, how stress changes genetic structure and affects life histories, how organisms cope with thermal stress through acclimation, and how environmental and genetic stress induce fluctuating asymmetry, shape selection pressure and cause extinction of populations. Finally, it discusses the role of stress in evolutionary change, from stress induced mutations and selection to speciation and evolution at the geological time scale. The book contains reviews and novel scientific results on the subject. It will be of interest to both researchers and graduate students and may serve as a text for graduate courses.