Developments over the past few years have revealed the remarkable versatility of RNA in any compartment of the cell, tasks that had been thought to be exclusively in the realm of proteins and even beyond. The chapters in this book written by leading investigators in the field provide insight into various promising avenues where RNA and nucleic acid derivatives including antisense RNAs, such as siRNA, miRNAs, amplification/selection (SELEX) generated aptamers as well as ribozymes are at the threshold of impacting medicine.

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.

A major success story of modem molecular biology is the development of technologies to clone and express specific genes. Current applications of recombinant gene products cover a wide spectrum, including gene therapy, production of bioactive pharmaceuticals, synthesis of novel biopolymers, agriculture and animal husbandry, and so on. Inherent in bringing these appli cations to fruition is the need to design "expression constructs" that will per mit the ready and specific detection and isolation of the defined recombinant gene products. Recombinant Protein Protocols grows out of the need for a laboratory manual on the detection and isolation of recombinantly expressed genes that covers both the background information and the practical laboratory recipes for these analyses. In this book, detailed and contemporary protocols are col lected to provide the reader with a wide-ranging number of methodologies to enhance the detection and isolation of their gene product(s) of interest. A large number of molecular tags and labels and their usage are described, including enzymes, ligand-binding moieties, immunodetectable molecules, as well as methods to detect interactive proteins, and gene expression-mediated alter ations in cellular activity. Chapters on in situ detection of gene expression deal with technologies that are currently being applied to the study of gene function and activity. Highlights of applications for recombinant gene expres sion technologies are provided to give readers exciting perspectives on the future of such technologies.

This proceedings is based on a joint meeting of the two IUFRO (International Union of Forestry Research Organizations) Working Parties, Somatic Cell Genetics (S2.04-07) and Molecular Genetics (S2.04-06) held in Gent, Belgium, 26-30 September, 1995. Although a joint meeting of the two Working Parties had been discussed in the past, this was the first such meeting that became a successful reality. In fact this meeting provided an excellent forum for discussions and interactions in forest bioteclUlology that encouraged the participants to vote for a next joint meeting. In the past decade rapid progress has been made in the somatic cell genetics and molecular genetics of forest trees. In order to cover recent developments in the broad area of biotechnology, the scientific program of the meeting was divided into several sessions. These included somatic embryogenesis, regeneration, transformation, gene expression, molecular markers, genome mapping, and biotic and abiotic stresses. The regeneration of plants, produced by organogenesis or somatic embryogenesis, is necessary not only for mass cloning of forest trees, but also for its application in genetic transformation and molecular biology. Although micropropagation has been achieved from juvenile tissues in a number of forest tree species, in vitro regeneration from mature trees remains a challenging problem in most hardwoods and conifers. The mechanisms involved in the transition from juvenile to mature phase in woody plants are poorly understood. This transition can now be investigated at the molecular level.

The effort to sequence the human genome is now moving toward a c- clusion. As all of the protein coding sequences are described, an increasing emphasis will be placed on understanding gene function and regulation. One important aspect of this analysis is the study of how transcription factors re- late transcriptional initiation by RNA polymerase II, which is responsible for transcribing nuclear genes encoding messenger RNAs. The initiation of Class II transcription is dependent upon transcription factors binding to DNA e- ments that include the core or basal promoter elements, proximal promoter elements, and distal enhancer elements. General initiation factors are involved in positioning RNA polymerase II on the core promoter, but the complex - teraction of these proteins and transcriptional activators binding to DNA e- ments outside the core promoter regulate the rate of transcriptional initiation. This initiation process appears to be a crucial step in the modulation of mRNA levels in response to developmental and environmental signals. Transcription Factor Protocols provides step-by-step procedures for key techniques that have been developed to study DNA sequences and the protein factors that regulate the transcription of protein encoding genes. This volume is aimed at providing researchers in the field with the well-detailed protocols that have been the hallmark of previous volumes of the Methods in Molecular (TM) Biology series.

Since its invention and subsequent development nearly 20 years ago, po- merase chain reaction (PCR) has been extensively utilized to identify numerous gene probes in vitro and in vivo. However, attempts to generate complete and full-length complementary cDNA libraries were, for the most part, fruitless and remained elusive until the last decade, when simple and rapid methods were developed. With current decoding and potential application of human genome information to genechips, there are urgent needs for identification of functional significance of these decoded gene sequences. Inherent in bringing these app- cations to fruition is the need to generate a complete and full-length cDNA library for potential functional assays of specific gene sequences. Generation of cDNA Libraries: Methods and Protocols serves as a laboratory manual on the evolution of generation of cDNA libraries, covering both ba- ground information and step-by-step practical laboratory recipes for which p- tocols, reagents, operational tips, instrumentation, and other requirements are detailed. The first chapter of the book is an overview of the basics of generating cDNA libraries, which include the following: (a) the definition of a cDNA library, (b) different kinds of cDNA libraries, (c) differences between methods for cDNA library generation using conventional approaches and novel stra- gies, including reverse generation of RNA repertoires from cDNA libraries, and (d) the quality of cDNA libraries.

Do our genes determine our behavior? Do education and environment have any influence at all? Do humans occupy a unique position in evolution? To clarify these provoking questions, the author takes the reader on an ambitious and entertaining journey through a variety of scientific disciplines. In doing so, he creates an image of human evolution that says that our entire individual knowledge is determined - to the smallest detail - by phylogeny. " ... before shocked humanists discard such radical theses as mere nonsense, they should not completely close their minds to the explanations of a biologist who says that we still know very little about the genetic determination of human behavior and that the invariance of many forms of behavior present in all cultures nourish the suspicion that the determining role of genes is probably far more comprehensive than we have ever dreamed."Wolfgang Wieser, translated from his review in "Merkur" (Sept./Oct. 1999)

The migration of stem cells has been found to be critical during early development for the organization of the embryonic body as well as during adult life with tissue homeostasis and regeneration of organ function. Due to the low frequency of these cells in vivo, problems in identifying and prospectively purifying tissue specific stem cells near homogeneity, and, most importantly, a lack of adequate technologies and protocols to study stem cell migration in vivo, this vital research has been quite difficult until recently. In "Stem Cell Migration: Methods and Protocols," experts in the field compile and highlight the standard and novel techniques that allow the studying of the migration of stem cells in one succinct manual. Including protocols on germ, neuronal, and hematopoietic stem cells, during development and adulthood with a clear emphasis on in vivo technologies, the volume also extends its coverage to in vitro approaches toward several developmentally-conserved signaling pathways. 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.

Practical and convenient, "Stem Cell Migration: Methods and Protocols" provides key, state-of the art information on experimental techniques for studying stem cell migration both at a cellular and molecular level in development, regeneration, and disease.

This book provides insights into the genetics and the latest advances in genomics research on the common bean, offering a timely overview of topics that are pertinent for future developments in legume genomics. The common bean (Phaseolus vulgaris L.) is the most important grain legume crop for food consumption worldwide, as well as a model for legume research, and the availability of the genome sequence has completely changed the paradigm of the ongoing research on the species. Key topics covered include the numerous genetic and genomic resources, available tools, the identified genes and quantitative trait locus (QTL) identified, and there is a particular emphasis on domestication. It is a valuable resource for students and researchers interested in the genetics and genomics of the common bean and legumes in general.

The staphylococci are important pathogenic bacteria responsible for a variety of diseases in humans and other animals. They are the most common cause of hospital-acquired infection. Antibiotic resistant strains (MRSA) have become endemic in hospitals in most countries, causing major public health issues. In addition, the incidence of new strains that cause severe community-acquired infections in healthy people is increasing and MRSA strains are emerging in agricultural and domestic animals. In the race to understand staphylococcal pathogenesis, the focus has been on genetics, as a bacterium can only do what its genes allow. The publication of the first staphylococcal whole genome sequence in 2001 paved the way for a greater understanding of the molecular basis of its virulence, evolution, epidemiology, and drug resistance. Since then, the available genomic data has mushroomed and this, coupled with the major advances in genetic know-how and the availability of better genetic tools, has

Although the field of quantitative genetics - the study of the genetic basis of variation in quantitative characteristics such as body size, or reproductive success - is almost 100 years old, its application to the study of evolutionary processes in wild populations has expanded greatly over the last few decades. During this time, the use of 'wild quantitative genetics' has provided insights into a range of important questions in evolutionary ecology, ranging from studies conducting research in well-established fields such as life-history theory, behavioural ecology and sexual selection, to others addressing relatively new issues such as populations' responses to climate change or the process of senescence in natural environments. Across these fields, there is increasing appreciation of the need to quantify the genetic - rather than just the phenotypic - basis and diversity of key traits, the genetic basis of the associations between traits, and the interaction between these genetic effects and the environment. This research activity has been fuelled by methodological advances in both molecular genetics and statistics, as well as by exciting results emerging from laboratory studies of evolutionary quantitative genetics, and the increasing availability of suitable long-term datasets collected in natural populations, especially in animals. Quantitative Genetics in the Wild is the first book to synthesize the current level of knowledge in this exciting and rapidly-expanding area. This comprehensive volume also offers exciting perspectives for future studies in emerging areas, including the application of quantitative genetics to plants or arthropods, unraveling the molecular basis of variation in quantitative traits, or estimating non-additive genetic variance. Since this book deals with many fundamental questions in evolutionary ecology, it should be of interest to graduate, post-graduate students, and academics from a wide array of fields such as animal behaviour, ecology, evolution, and genetics.

The discovery of DNA as the genetic material brought great hope to scientists all over the world. It was believed that many of the lingering questions in genetics and the mechanisms of heredity would fnally be answered. However, as often is the case in science, more qu- tions arose out of this discovery. What defnes a gene? What are the mechanisms of gene regulation? Further discovery and technological innovations brought about sequencing techniques that allowed the study of complete genomes from many organisms, including Arabidopsis and humans. Despite all the excitement surrounding these technologies, many features of the genome remained unclear. Peculiar characteristics in genome composition such as signifcant redundancy consisting of many repetitive elements and noncoding sequences, active transcriptional units with no protein product, and unusual sequences in promoter regions added to the mysteries of genetic make-up and gene regulation. Indeed, the more we discovered about the genome, the more diffcult it became to understand the complexity of cellular function and regulation. Out of the study of the intricacies of the genome and gene regulation, arose a new science that was independent of actual DNA changes, but critical in maintaining gene regulation and genetic stability. Epigenetics, literally translated as "above genetics," is the science that describes the mechanisms of heritable changes in gene regulation that does not involve modifcations of DNA sequence. These changes may last through somatic cell division and, in some cases, throughout multiple generations.

This volume collects a series of protocols describing the kinds of infrastructures, training, and standard operating procedures currently available to actualize the potential of stem cells for regenerative therapies. Stem Cells and Good Manufacturing Practices: Methods, Protocols, and Regulations pulls together key GMP techniques from laboratories around the world. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Inclusive and authoritative, Stem Cells and Good Manufacturing Practices: Methods, Protocols, and Regulations will be an invaluable resource to both basic and clinical practitioners in stem cell biology.

Microarray technology is a major experimental tool for functional genomic explorations, and will continue to be a major tool throughout this decade and beyond. The recent explosion of this technology threatens to overwhelm the scientific community with massive quantities of data. Because microarray data analysis is an emerging field, very few analytical models currently exist. Methods of Microarray Data Analysis is one of the first books dedicated to this exciting new field. In a single reference, readers can learn about the most up-to-date methods ranging from data normalization, feature selection and discriminative analysis to machine learning techniques. Currently, there are no standard procedures for the design and analysis of microarray experiments. Methods of Microarray Data Analysis focuses on two well-known data sets, using a different method of analysis in each chapter. Real examples expose the strengths and weaknesses of each method for a given situation, aimed at helping readers choose appropriate protocols and utilize them for their own data set. In addition, web links are provided to the programs and tools discussed in several chapters. This book is an excellent reference not only for academic and industrial researchers, but also for core bioinformatics/genomics courses in undergraduate and graduate programs.

This book presents a novel molecular description for understanding the regulatory mechanisms behind the autonomy and self-organization in biological systems. Chapters focus on defining and explaining the regulatory molecular mechanisms behind different aspects of autonomy and self-organization in the sense of autonomous coding, data processing, structure (mass) formation and energy production in a biological system. Subsequent chapters discuss the cross-talk among mechanisms of energy, and mass and information, transformation in biological systems. Other chapters focus on applications regarding therapeutic approaches in regenerative medicine. Molecular Mechanisms of Autonomy in Biological Systems is an indispensable resource for scientists and researchers in regenerative medicine, stem cell biology, molecular biology, tissue engineering, developmental biology, biochemistry, biophysics, bioinformatics, as well as big data sciences, complexity and soft computing.

By the end of the 1980s only two microtubule-dependent motors, the plus end-directed kinesin and the minus end-directed cytoplasmic dynein, had been identified. At the time, these two motors seemed almost sufficient to explain directional motility events on polar microtubule tracks in the cell. No- theless, shortly after, the tip of the iceberg began to emerge with the identi- cation of proteins containing in their sequences a domain found in kinesin. This domain, called the "motor domain," conferred on these proteins the essential property of moving on microtubules, using the energy derived from ATP hydro- sis. Since then, the identification of new proteins belonging to the kinesin superfamily of microtubule-dependent motors has gone at such a pace that nowadays more than 200 entries with motor domain sequences are deposited in the database. Kinesin family members are found in all eukaryotic org- isms tested. They present a wide range of domain organizations with a motor domain located at different positions in the molecule. Their motility prop- ties are also variable in directionality, velocity, and such other characteristics as bundling activity and processivity. Finally, and most important, they p- ticipate in a multitude of cellular functions. Our understanding of many cel- lar events, such as mitotic spindle assembly and neuronal transport, to cite only two, has progressed substantially in the last few years thanks to the id- tification of these motors.

Epigenetic Regulation of Cancer in Response to Chemotherapy, Volume 158 of the Advances in Cancer Research series, highlights new advances in the field, with this new volume presenting interesting chapters. Each chapter is written by an international board of authors.

Genetic erosions in plant cell cultures, especially in chromosome number and ploidy level, have now been known for over 25 years. Until the mid -1970ssuch changes were consideredundesirable and thereforediscarded because the main emphasis wason clonal propagation and genetic stability of cultures. However, since the publication on somaclonal variation by Larkin and Scowcroft (1981) there has been a renewed interest to utilize these in vitro obtained variations for crop improvement. Studies conduc- ted during the last decade have shown that callus cultures, especially on peridical subculturing over an extended period of time, undergo morpho- logical and genetic changes, i. e. polyploidy, aneuploidy, chromosome breakage, deletions, translocations, gene amplification, inversions, muta- tions, etc. In addition, there are changes at the molecular and biochemical levelsincluding changes in the DNA, enzymes,proteins, etc. Suchchanges are now intentionally induced, and useful variants are selected. For instance in agricultural crops such as potato, tomato, tobacco, maize, rice and sugarcane, plants showing tolerance to a number of diseases, viruses, herbicides and salinity, have been isolated in cell cultures. Likewise induction of male sterility in rice, and wheat showing various levels of fer- tility and gliadin, have been developed in vitro. These academic excercises open new avenues for plant breeders and pathologists. Another area of tremendous commercial importance in the pharmaceuti- cal industry is the selection of cell lines showing high levels of medicinal and industrial compounds. Already high shikonin containing somaclones in Lithospermum are being used commercially.

Developmental Instability: Its Origins and Evolutionary Implications is a collection of papers and transcribed discussions from a conference held in Tempe, Arizona in June 1993. The papers represent a wide range of contributions, from the empirical to the theoretical, and include methods for measuring developmental instability across a variety of taxa and traits. This volume presents contrasting views on how to assess developmental instability as well as on the relationship of instability to genotypic factors, environmental factors and the action of natural and sexual selection. Readers will derive a working knowledge of the best way to assess developmental instability and will be able to design future work in an authoritative way.

Bioinformatics is an integrative field of computer science, genetics, genomics, proteomics, and statistics, which has undoubtedly revolutionized the study of biology and medicine in past decades. It mainly assists in modeling, predicting and interpreting large multidimensional biological data by utilizing advanced computational methods. Despite its enormous potential, bioinformatics is not widely integrated into the academic curriculum as most life science students and researchers are still not equipped with the necessary knowledge to take advantage of this powerful tool. Hence, the primary purpose of our book is to supplement this unmet need by providing an easily accessible platform for students and researchers starting their career in life sciences. This book aims to avoid sophisticated computational algorithms and programming. Instead, it will mostly focus on simple DIY analysis and interpretation of biological data with personal computers. Our belief is that once the beginners acquire these basic skillsets, they will be able to handle most of the bioinformatics tools for their research work and to better understand their experimental outcomes. The third volume is titled In Silico Life Sciences: Agriculture. It focuses on plant genetic, genomic, transcriptomic, proteomic and metabolomics data. Using examples of new crop diseases-emergence, crop productivity and biotic/abiotic stress tolerance, this book illustrates how bioinformatics can be an integral components of modern day plant science research.

Leishmania is a vector-borne pathogenic parasite found in 88 countries worldwide and is the causative agent of leishmaniasis. The different Leishmania species infect macrophages and dendritic cells of the host immune system, causing symptoms that range from disfiguring cutaneous and mucocutaneous lesions, widespread destruction of mucous membranes, or visceral disease affecting the haemopoetic organs. The recent publication of the complete genome sequences of three different Leishmania species provides new insights into this leading pathogen and presents scientists with an exciting resource to improve the understanding of its complex molecular and cellular biology. In this book, internationally recognized Leishmania experts critically review the most important aspects of current Leishmania research, providing the first coherent picture of the organism's molecular and cellular biology since the publication of the genome sequence. Chapters are written from a molecular and genomic perspective and discuss in depth Leishmania-specific aspects of trypanosomatid biology and pathology. Topics include: diagnosis and epidemiology, genome structure and content, regulation of gene expression, the Leishmania proteome, the Leishmania metabolome, Leishmania differentiation, interaction with the sand fly vector, drug discovery, drug resistance, and much more. This will be essential reading for all researchers working with Leishmania, trypanosomes, and protozoa; and is recommended for all biology and medical libraries.

This book aims at defining and reassessing the role of population genetics in conservation biology and seeks to identify the progress made in the field during the last decade. It deals with conservation genetics from several currently researched points of view, namely, ecological and demographic measures of rarity or population persistence, loss of genetic variation, inbreeding, reduced migration rates and increased selective pressures under stress and the role of social behaviour and metapopulation structure. The use of molecular variation as the basis of naming or selecting target taxa and some strategic decisions about genetic variance maximization in the conserved population or community units are analysed. Several case studies and scenarios illustrate the application of genetic information in conservation practices.

These are indeed exciting times to be a microbiologist. With one of the buzzwords of the past decade-"Biodiversity," and microbes are reveling in the attention as they represent by far most of the biodiversity on Earth. Microbes can thrive in almost any environment where there is an exploitable energy source, and, as a result, the possible existence of microbial life elsewhere in the solar system has stimulated the imaginations of many. Extremophiles have taken center stage in these investigations, and thermophiles have taken on the lead roles. Consequently, in the past decade there has been a surge of interest and research in the Ecology, Biology, and Biotechnology of microorganisms from thermal environments. Many of the foundations of thermophile research were laid in Yellowstone National Park, primarily by the research of Professor Thomas Brock's laboratory in the late 1960s and early 1970s. The upper temperature for life was debated, the first thermophilic archeum discovered (although it was only later shown to be an archeum by ribosomal cataloging), and the extremes of light, temperature, pH on the physiology of microorga nisms were explored. Interest in thermophiles increased steadily in the 1970s, and with the discovery of deep-sea hydrothermal vents in 1977, thermophilic research began its expo nential explosion. The development of Taq polymerase in the polymerase chain reaction (peR) focused interest on the biotechnological potential of thermophilic microorganisms and on the thermal features in Yellowstone National Park."