This volume is designed to be a resource of proven techniques and approaches for probing the activities of bacterial, eukaryotic, and archaeal RNA polymerases. This book features a collection of in vitro and in vivo technologies that will permit researchers to purify and probe the position and stability of RNA polymerase complexes at different points of the transcription cycle, analyze the various translocations and intermolecular movements associated with catalysis, define recruitment strategies, probe the roles of transcription factors in each stage of the cycle, highlight conserved and disparate fidelity mechanisms, analyze the resultant transcripts, and study coordination of the nascent mRNA synthesis by the RNA polymerase and mRNA translation by the ribosome. 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. Practical and timely, Bacterial Transcriptional Controls: Methods and Protocols highlights the breadth and depth of techniques that are likely to continue shaping the transcription community in the future.

From Dolly the sheep to Frankenfood, life-saving medicines, and beyond, this insightful work describes the technology and controversy behind genetic engineering. From the publication of Aldous Huxley's Brave New World in 1932 to the cloning of Dolly the sheep in 1996, the public has long been fascinated by the idea that humans may one day be able to mold or even create life. In less than 30 years, genetic engineering has itself mutated from science fiction to science fact. Supporters claim such innovations as genetically modified crops and gene therapy are poised to bring unparalleled benefits by eliminating hunger and hereditary disease, whereas critics warn the dream could easily become a nightmare. Packed with key facts and analysis, Genetic Engineering: A Reference Handbook, Second Edition provides an expert guide to the very latest discoveries in genetic engineering and genetic modification and the technology's complex ethical, scientific, and economic implications. Includes an expert guide to print and online resources on genetic engineering and related areas Features a comprehensive glossary designed for the general reader

Classical methods for microbial strain engineering, used to improve the production of bioproducts, have serious drawbacks and have been found to be unsuitable for complex strain development applications. In Strain Engineering: Methods and Protocols, powerful new genetic engineering-based strain engineering methods are presented for rational modification of a variety of model organisms. These methods are particularly powerful when utilized to manipulate microbes for which sequenced and annotated genomes are available. Collectively, these methods systematically introduce genome alterations in a precise manner, allowing the creation of novel strains carrying only desired genome alterations. In the first section, E. coli-based bacterial strain engineering strategies are reviewed, while the second section presents analogous microbial engineering strategies for eukaryotic cells using the yeast Saccharomyces cerevisiae as a model. The third section covers examples of the proliferative adaptations of these base technologies to strain engineer industrially important prokaryotic or eukaryotic microbial systems. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and accessible, Strain Engineering: Methods and Protocols serves as an ideal guide to scientists in academia, pharmaceutical science, and biotechnology who perform microbial strain engineering.

The rapid progression of genetics and molecular biology has turned chromosomal engineering from science fiction to reality, with the successful production of transgenic animals with engineered chromosomes and chromosomes developed for pharmaceutical protein production which are now ready for the medical industry. Mammalian Chromosome Engineering: Methods and Protocols provides the reader with up-to date information on this rapidly evolving field and strives to take the reader into the exciting realm of chromosomal engineering from the basic principles to the practical applications of these new technologies. The five overview and ten protocol chapters cover the engineering of chromosomes with extrachromosomal vectors and transposon systems, the manipulation of naturally occurred minichromosomes, the generation and engineering of synthetic artificial chromosomes, and the induced de novo platform artificial chromosome system. Written in the highly successful Methods in Molecular Biology (TM) series format, protocols chapters contain brief 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, Mammalian Chromosome Engineering: Methods and Protocols serves as a bench-side resource for current protocols and aims to help scientists to explore the many prospects for future research and vital applications.

Plant molecular biology came to the fore in the early 1980s and there has been tremendous growth in the subject since then. The study of plant genes and genomes and the development of techniques for the incorporation of novel or modified genes into plants eventually led to the commercialisation of genetically modified (GM) crops in the mid-1990s. This was seen as the start of a biotechnological revolution in plant breeding. However, plant biotechnology has become one of the hottest debates of the age and, in Europe at least, one of the greatest challenges that plant scientists have ever faced.This book covers the history and development of the science and techniques that underpin plant biotechnology. It describes the GM crops that are or have been grown commercially around the world, including failures as well as successes, and the new varieties that are being developed. The safety record of GM crops is reviewed together with the legislation that has been adopted to cover their use. The book also deals with the concerns of consumers, the GM crop debate and the prospects for the technology. In the second edition, sections on current GM crops and future developments in plant biotechnology have been greatly expanded, while those on techniques, legislation and the GM crop debate have also been updated.The book is a concise, comprehensive and readable study that is accessible to a general readership with a scientific background but also provides useful information for the specialist.

Arguing that human evolution has come to a stand-still, this book sets out to explore the evolutionary steps that have defined life on this planet. It describes the stages from cosmic to chemical and biological evolution and to the development of civilization and culture. From this scientific approach, the author concludes that new rules of ethics are required in order to maintain and improve the civilization and culture of humanity. Matare argues for corrective genetic interference and explores the moral implications.

This thesis reports on the development of a fully integrated and automated microsystem consisting of low-cost, disposable plastic chips for DNA extraction and PCR amplification, combined with a reusable glass capillary array electrophoresis chip, which can be employed in a modular-based format for genetic analysis. In the thesis, DNA extraction is performed by adopting a filter paper-based method, followed by an "in-situ" PCR carried out directly in the same reaction chamber of the chip without elution. PCR products are then co-injected with sizing standards into separation channels for detection using a novel injection electrode. The entire process is automatically carried out by a custom-made compact control and detection instrument. The author thoroughly tests the system's performance and reliability by conducting rapid genetic screening of mutations on congenital hearing loss and pharmacogenetic typing of multiple warfarin-related single-nucleotide polymorphisms. The successful development and operation of this microsystem establishes the feasibility of rapid "sample-in-answer-out" testing in routine clinical practice.

During the past 15 years, cellular and molecular approaches have emerged as valuable adjuncts to supplement and complement conventional breeding methods for a wide variety of crop plants. Biotechnology increasingly plays a role in the creation, conservation, characterization and utilization of genetic variability for germplasm enhancement. For instance, anther/microspore culture, somaclonal variation, embryo culture and somatic hybridization are being exploited for obtaining incremental improvement in the existing cultivars. In addition, genes that confer insect- and disease-resistance, abiotic stress tolerance, herbicide tolerance and quality traits have been isolated and re-introduced into otherwise sensitive or susceptible species by a variety of transgenic techniques. Together these transformative methodologies grant access to a greater repertoire of genetic diversity as the gene(s) may come from viruses, bacteria, fungi, insects, animals, human beings, unrelated plants or even be artificially derived. Remarkable achievements have been made in the production, characterization, field evaluation and commercialization of transgenic crop varieties worldwide. Likewise, significant advances have been made towards increasing crop yields, improving nutritional quality, enabling crops to be raised under adverse conditions and developing resistance to pests and diseases for sustaining global food and nutritional security. The overarching purpose of this 3-volume work is to summarize the history of crop improvement from a technological perspective but to do so with a forward outlook on further advancement and adaptability to a changing world. Our carefully chosen "case studies of important plant crops" intend to serve a diverse spectrum of audience looking for the right tools to tackle complicated local and global issues.

Appropriate for a wide range of disciplines, from biology to non-biology, law and nursing majors, "DNA and Biotechnology" uses a straightforward and comprehensive writing style that gives the educated layperson a survey of DNA by presenting a brief history of genetics, a clear outline of techniques that are in use, and highlights of breakthroughs in hot topic scientific discoveries.

Engaging and straightforward scientific writing style Comprehensive forensics chapter Parallel Pedagogic material designed to help both readers and teachers. Highlights in the latest scientific discoveries Outstanding full-color illustration that walk reader through complex concepts

Due to the possibility that petroleum supplies will be exhausted in the next decades to come, more and more attention has been paid to the production of bacterial pl- tics including polyhydroxyalkanoates (PHA), polylactic acid (PLA), poly(butylene succinate) (PBS), biopolyethylene (PE), poly(trimethylene terephthalate) (PTT), and poly(p-phenylene) (PPP). These are well-studied polymers containing at least one monomer synthesized via bacterial transformation. Among them, PHA, PLA and PBS are well known for their biodegradability, whereas PE, PTT and PPP are probably less biodegradable or are less studied in terms of their biodegradability. Over the past years, their properties and appli- tions have been studied in detail and products have been developed. Physical and chemical modifications to reduce their cost or to improve their properties have been conducted. PHA is the only biopolyester family completely synthesized by biological means. They have been investigated by microbiologists, molecular biologists, b- chemists, chemical engineers, chemists, polymer experts, and medical researchers for many years. PHA applications as bioplastics, fine chemicals, implant biomate- als, medicines, and biofuels have been developed. Companies have been est- lished for or involved in PHA related R&D as well as large scale production. It has become clear that PHA and its related technologies form an industrial value chain in fermentation, materials, feeds, and energy to medical fields.

This book provides exhaustive information on several recent technologies that are employed for sugarcane improvement through biotechnology and will be of great interest to plant scientists, biotechnologists, molecular biologists and breeders who work on sugarcane crop. Topics discussed in this volume include genomics and transcriptomics, transgenic sugarcane for trait improvement, potential candidate promoters, new strategies for transformation, molecular farming, sugarcane as biofuel, chloroplast transformation, and genome editing.

DNA replication is arguably the most crucial process at work in living cells. It is the mechanism by which organisms pass their genetic information from one generation to the next and life on Earth would be unthinkable without it. Despite the discovery of DNA structure in the 1950s, the mechanism of its replication remains rather elusive. This work makes important contributions to this line of research. In particular, it addresses two key questions in the area of DNA replication: which evolutionary forces drive the positioning of replication origins in the chromosome and how is the spatial organization of replication factories achieved inside the nucleus of a cell?. A cross-disciplinary approach uniting physics and biology is at the heart of this research. Along with experimental support, statistical physics theory produces optimal origin positions and provides a model for replication fork assembly in yeast. Advances made here can potentially further our understanding of disease mechanisms such as the abnormal replication in cancer.

Rapid changes and significant progress have been made in the Agrobacterium field, such as genetically transforming plants for both basic research purposes and agricultural development. In Agrobacterium Protocols, Third Edition, Volumes 1 and 2, a team of leading experts and veteran researchers describe in detail techniques for delivering DNA to plant cells and permanently altering their genomes. This edition emphasizes agricultural crops and plant species with economic values, with updated protocols on 32 plant species and protocols involving 19 new species. Together with the 1st and 2nd editions, these two volumes offer Agrobacterium-mediated genetic transformation protocols for a total of 76 plant species. For a number of important plants such as rice, barley, wheat and citrus, multiple protocols using different starting plant materials for transformation are included. Volume 2 contains 29 chapters with updated techniques for industrial plants, root plants, nuts and fruits, tropic plants, and other important plant species. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Agrobacterium Protocols, Third Edition facilitates the transfer of this rapidly developing technology to all researchers in both fundamental and applied biology.

This timely volume explores the use of CRISPR-Cas9 for genome editing, presenting cutting-edge techniques and their applications in treatment of disease. The chapters describe latest methods such as use of targetable nucleases, investigation of the non-coding genome, mouse genome editing, increasing of knock-in efficiency in mouse zygotes, and generation of reporter stem cells; the text contextualizes these methods in treatment of cardiovascular disease, diabetes mellitus, retinitis pigmentosa, and others. The final chapters round out the book with a discussion of controversies and future directions. Genome Editing is an essential, of-the-moment contribution to this rapidly growing field. Drawing from a wealth of international perspectives, it presents novel techniques and applications for the engineering of the human genome. This book is essential reading for all clinicians and researchers in stem cells, regenerative medicine, genomics, biochemical and biomedical engineering- especially those interested in learning more about genome editing and applying it in a targeted, specific way.

The purpose of this publication is to elucidate the biological aspect of the abiotic stress response from the field to the molecular level in horticultural plants. This book is unique in that it concerns the basic aspect of abiotic stress biology and research progress at the molecular level in model plants or major field crops, as it focuses mainly on the abiotic stress response in existing horticultural plants. Many readers interested in plant abiotic stress biology are aware of the application of the latest findings to agricultural production, and this book will have a special appeal for those readers. The book will be of interest to scientists and graduate students who are involved in the research, development, production, processing, and marketing of horticultural products, including those in developing countries who are interested in high tech and advanced science in this field. The application of the latest findings to agricultural production is particularly useful. Stress tolerance mechanisms in horticultural crops are gaining importance, because most agricultural regions are predicted to experience considerably more extreme environmental fluctuations due to global climate change. Further, because of recent progress in next-generation sequencing technologies, the postgenomic era is impending not only in model plants and major cereal crops but also in horticultural crops, which comprise a great diversity of species. This book provides information on the physiological aspects of the abiotic stress response in horticultural plants, which is considered essential for postgenomic research.

Plant tissue culture and advanced biotechnologies have proven to be influential tools that complement conventional breeding and accelerate development of many medicinal plants. Various approaches, such as pathway engineering, precursor feeding, transformation, elicitation with biotic and abiotic elicitors and scaling up in bioreactors, have been explored to improve the production of secondary metabolites from different medicinal plants. This book provides a comprehensive description of various studies, carried out on in vitro culture and hairy root cultures of Catharanthus roseus, Silybum marianum and Digitalis species which have been considered as alternative sources for the production of anti-tumour compounds, flavonolignans and cardenolides. Specific focus is on elicitation strategy for increasing production of bioactive compounds of C. roseus L., S. marianum and Digitalis species to overcome the constrains of conventional propagation. This book is valuable for researchers or students working on medicinal plants, phytochemistry, and plant tissue culture. It also serves as a reference for the pharmaceutical industry.

th We compiled this volume mostly from presentations at the 6 International Plant Cold Hardiness Seminar (PCHS) after consulting with Professor Tony H. H. Chen, Oregon State University, USA, Professor Pekka Heino, University of Helsinki, Finland, th and Dr. Gareth J. Warren, University of London, Surrey, UK. The 6 International PCHS was held at the Unitas Congress Center, Helsinki, Finland from July 1-5, 2001. There were 110 registered scientists at the serttinar representing 20 countries: Australia, Belgium, Canada, Chile, the Czech Republic, Denmark, Estonia, Finland, Gennany, Hungary, Iceland, Italy, Japan, Norway, Poland, Spain, Sweden, Taiwan, United Kingdom, and United States of America. The infonnation compiled represents the state of the art of research in phmt cold hardiness in tenns of gene regulation, gene expression, signal transduction, the physiology of cold hardiness and, ultimately, the genetic engineering for cold tolerant plants. The International PCHS was initiated in 1977 at the University of Minnesota, St. Paul, Minnesota. It has been traditionally held at 5-year intervals at various locations. th Because of the rapid advances of research in plant cold hardiness, attendees at the 6 meeting unanimously adopted a resolution to hold the seminar in 3-year intervals instead of 5 in the future. Consequently, the next seminar will be held in 2004 in Sapporo, Japan, and Professor Seizo Fujikawa from Hokkaido University will serve as the host.

Drawing on a wide range of interviews and primary and secondary sources, this book investigates the dynamic interactions between national regulatory formation and the global biopolitics of regenerative medicine and human embryonic stem cell science.

Genetic Engineering, Volume 24 contains discussions of contemporary and relevant topics in genetics, including:

-Gene silencing: principles and applications,
-Integrins and the myocardium,
-Plant virus gene vectors: biotechnology and applications in agriculture and medicine,
-Novel approaches to controlling transcription,
-Use of DNA polymorphisms in genetic mapping,
-Application of FLP/FRT site-specific DNA recombination system in plants.
This principles and methods approach to genetics and genetic engineering is essential reading for all academics, bench scientists, and industry professionals wishing to take advantage of the latest and greatest in this continuously emerging field.

With an increasing human population and a decreasing amount of arable land, creative improvements in agriculture will be a necessity in the coming decades to maintain or improve the standard of living. In Plant Chromosome Engineering: Methods and Protocols, expert researchers present techniques for the modification of crops and other plant species in order to achieve the goal of developing the much needed novel approaches to the production of food, feed, fuel, fiber, and pharmaceuticals. This volume examines vital topics such as transformation procedures, chromosome painting, production of engineered minichromosomes, gene targeting and mutagenesis, site specific integration, gene silencing, protein expression, chromosome sorting and analysis, protocols for generating chromosomal rearrangements, enhancer trapping, and means of studying chromosomes in vivo. As a part of the highly successful Methods in Molecular Biology(TM) series, the methodological chapters include brief introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and professional tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Plant Chromosome Engineering: Methods and Protocols highlights the spectrum of tools currently available for modifying plant genomes and chromosomes and provides the foundation for crucial future developments.

Genetic Engineering, Volume 25 contains discussions of contemporary and relevant topics in genetics, including:
- Genotyping by Mass Spectrometry;
- Development of Targeted Viral Vectors for Cardiovascular Gene Therapy;
- Practical Applications of Rolling Circle Amplification of DNA Templates;
- Bacterial ION Channels;
- Applications of Plant Antiviral Proteins;
- The Bacterial Scaffoldin: Structure, Function and Potential Applications in the Nanosciences. This principles and methods approach to genetics and genetic engineering is essential reading for all academics, bench scientists, and industry professionals wishing to take advantage of the latest and greatest in this continuously emerging field.

Aspects of genetic engineering research emphasized in this volume are applications to plants (crop plants and grass, both important for human needs) and new methodologies, such as Tar cloning, which make it much easier to isolate specific regions from complex genomes. Another subject discussed is linear DNA replication of prokaryotes.