Production of Recombinant Proteins

Production of Recombinant Proteins
Author: Gerd Gellissen
Publisher: John Wiley & Sons
Total Pages: 429
Release: 2006-03-06
Genre: Science
ISBN: 3527604413

While the choices of microbial and eukaryotic expression systems for production of recombinant proteins are many, most researchers in academic and industrial settings do not have ready access to pertinent biological and technical information since it is normally scattered throughout the scientific literature. This book closes the gap by providing information on the general biology of the host organism, a description of the expression platform, a methodological section -- with strains, genetic elements, vectors and special methods, where applicable -- as well as examples of proteins produced with the respective platform. The systems thus described are well balanced by the inclusion of three prokaryotes (two Gram-negatives and one Gram-positive), four yeasts, two filamentous fungi and two higher eukaryotic cell systems -- mammalian and plant cells. Throughout, the book provides valuable practical and theoretical information on the criteria and schemes for selecting the appropriate expression platform, the possibility and practicality of a universal expression vector, and on comparative industrial-scale fermentation, with the production of a recombinant Hepatitis B vaccine chosen as an industrial example. With a foreword by Herbert P. Schweizer, Colorado State University, USA: "As a whole, this book is a valuable and overdue resource for a varied audience. It is a practical guide for academic and industrial researchers who are confronted with the design of the most suitable expression platform for their favorite protein for technical or pharmaceutical purposes. In addition, the book is also a valuable study resource for professors and students in the fields of applied biology and biotechnology."

Gene Expression Systems

Gene Expression Systems
Author: Joseph M. Fernandez
Publisher: Academic Press
Total Pages: 504
Release: 1999
Genre: Science
ISBN:

Recombinant gene expression is the fastest growing area in the study of molecular biology. By the time the Human Genome Project is completed (~2002), several thousand sequences will be known, but the purpose of the resultant expression products will remain a mystery. Gene discovery requires efficient expression systems for determining the structure and function of gene products. Gene Expression Systems covers a variety of promoters and host organisms that researchers can tailor to their specific needs.

What the Face Reveals

What the Face Reveals
Author: Paul Ekman
Publisher: Oxford University Press
Total Pages: 663
Release: 2005-04-14
Genre: Psychology
ISBN: 0199792720

While we have known for centuries that facial expressions can reveal what people are thinking and feeling, it is only recently that the face has been studied scientifically for what it can tell us about internal states, social behavior, and psychopathology. Today's widely available, sophisticated measuring systems have allowed us to conduct a wealth of new research on facial behavior that has contributed enormously to our understanding of the relationship between facial expression and human psychology. The chapters in this volume present the state-of-the-art in this research. They address key topics and questions, such as the dynamic and morphological differences between voluntary and involuntary expressions, the relationship between what people show on their faces and what they say they feel, whether it is possible to use facial behavior to draw distinctions among psychiatric populations, and how far research on automating facial measurement has progressed. The book also includes follow-up commentary on all of the original research presented and a concluding integration and critique of all the contributions made by Paul Ekman. As an essential reference for all those working in the area of facial analysis and expression, this volume will be indispensable for a wide range of professionals and students in the fields of psychology, psychiatry, and behavioral medicine.

Recombinant protein expression in microbial systems

Recombinant protein expression in microbial systems
Author: Eduardo A. Ceccarelli
Publisher: Frontiers E-books
Total Pages: 103
Release: 2014-10-02
Genre: Biotechnology
ISBN: 2889192946

With the advent of recombinant DNA technology, expressing heterologous proteins in microorganisms rapidly became the method of choice for their production at laboratory and industrial scale. Bacteria, yeasts and other hosts can be grown to high biomass levels efficiently and inexpensively. Obtaining high yields of recombinant proteins from this material was only feasible thanks to constant research on microbial genetics and physiology that led to novel strains, plasmids and cultivation strategies. Despite the spectacular expansion of the field, there is still much room for progress. Improving the levels of expression and the solubility of a recombinant protein can be quite challenging. Accumulation of the product in the cell can lead to stress responses which affect cell growth. Buildup of insoluble and biologically inactive aggregates (inclusion bodies) lowers the yield of production. This is particularly true for obtaining membrane proteins or high-molecular weight and multi-domain proteins. Also, obtaining eukaryotic proteins in a prokaryotic background (for example, plant or animal proteins in bacteria) results in a product that lack post-translational modifications, often required for functionality. Changing to a eukaryotic host (yeasts or filamentous fungi) may not be a proper solution since the pattern of sugar modifications is different than in higher eukaryotes. Still, many advances in the last couple of decades have provided to researchers a wide variety of strategies to maximize the production of their recombinant protein of choice. Everything starts with the careful selection of the host. Be it bacteria or yeast, a broad list of strains is available for overcoming codon use bias, incorrect disulfide bond formation, protein toxicity and lack of post-translational modifications. Also, a huge catalog of plasmids allows choosing for different fusion partners for improving solubility, protein secretion, chaperone co-expression, antibiotic resistance and promoter strength. Next, controlling culture conditions like temperature, inducer and media composition can bolster recombinant protein production. With this Research Topic, we aim to provide an encyclopedic account of the existing approaches to the expression of recombinant proteins in microorganisms, highlight recent discoveries and analyze the future prospects of this exciting and ever-growing field.

Cell-Free Protein Expression

Cell-Free Protein Expression
Author: James R. Swartz
Publisher: Springer Science & Business Media
Total Pages: 213
Release: 2012-12-06
Genre: Science
ISBN: 3642593372

Cell-free protein synthesis is coming of age! Motivated by an escalating need for efficient protein synthesis and empowered by readily accessible cell-free protein synthesis kits, the technology is expanding both in the range of feasible proteins and in the ways that proteins can be labeled and modified. This volume follows "Cell-Free Translation Systems", edited by Professor Alexander S. Spirin in 2002. Since then, an impressive collection of new work has emerged that demonstrates a substantial expansion of capability. In this volume, we show that proteins now can be efficiently produced using PCR products as DNA templates and that even membrane proteins and proteins with multiple disulfide proteins are obtained at high yields. Many additional advances are also presented. It is an exciting time for protein synthesis technology.

Industrial Enzyme Applications

Industrial Enzyme Applications
Author: Andreas Vogel
Publisher: John Wiley & Sons
Total Pages: 430
Release: 2019-10-28
Genre: Science
ISBN: 3527343857

This reference is a "must-read": It explains how an effective and economically viable enzymatic process in industry is developed and presents numerous successful examples which underline the efficiency of biocatalysis.

The Architectural Expression of Environmental Control Systems

The Architectural Expression of Environmental Control Systems
Author: George Baird
Publisher: Taylor & Francis
Total Pages: 287
Release: 2003-09-02
Genre: Architecture
ISBN: 1135920524

Examines the ways design teams can take a more creative approach to the design and expression of both active and passive environmental control systems. Case Studies from around the world, including many award-winning buildings.

Cell-Free Gene Expression

Cell-Free Gene Expression
Author: Ashty S. Karim
Publisher: Humana
Total Pages: 435
Release: 2022-01-06
Genre: Science
ISBN: 9781071619971

This detailed volume explores perspectives and methods using cell-free expression (CFE) to enable next-generation synthetic biology applications. The first section focuses on tools for CFE systems, including a primer on DNA handling and reproducibility, as well as methods for cell extract preparation from diverse organisms and enabling high-throughput cell-free experimentation. The second section provides an array of applications for CFE systems, such as metabolic engineering, membrane-based and encapsulated CFE, cell-free sensing and detection, and educational kits. Written for the highly successful Methods in Molecular Biology series, 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 practical, Cell‐Free Gene Expression: Methods and Protocols serves as an ideal guide for researchers seeking technical methods to current aspects of CFE and related applications.

Stress and Environmental Regulation of Gene Expression and Adaptation in Bacteria

Stress and Environmental Regulation of Gene Expression and Adaptation in Bacteria
Author: Frans J. de Bruijn
Publisher: John Wiley & Sons
Total Pages: 1472
Release: 2016-07-13
Genre: Science
ISBN: 1119004896

Bacteria in various habitats are subject to continuously changing environmental conditions, such as nutrient deprivation, heat and cold stress, UV radiation, oxidative stress, dessication, acid stress, nitrosative stress, cell envelope stress, heavy metal exposure, osmotic stress, and others. In order to survive, they have to respond to these conditions by adapting their physiology through sometimes drastic changes in gene expression. In addition they may adapt by changing their morphology, forming biofilms, fruiting bodies or spores, filaments, Viable But Not Culturable (VBNC) cells or moving away from stress compounds via chemotaxis. Changes in gene expression constitute the main component of the bacterial response to stress and environmental changes, and involve a myriad of different mechanisms, including (alternative) sigma factors, bi- or tri-component regulatory systems, small non-coding RNA’s, chaperones, CHRIS-Cas systems, DNA repair, toxin-antitoxin systems, the stringent response, efflux pumps, alarmones, and modulation of the cell envelope or membranes, to name a few. Many regulatory elements are conserved in different bacteria; however there are endless variations on the theme and novel elements of gene regulation in bacteria inhabiting particular environments are constantly being discovered. Especially in (pathogenic) bacteria colonizing the human body a plethora of bacterial responses to innate stresses such as pH, reactive nitrogen and oxygen species and antibiotic stress are being described. An attempt is made to not only cover model systems but give a broad overview of the stress-responsive regulatory systems in a variety of bacteria, including medically important bacteria, where elucidation of certain aspects of these systems could lead to treatment strategies of the pathogens. Many of the regulatory systems being uncovered are specific, but there is also considerable “cross-talk” between different circuits. Stress and Environmental Regulation of Gene Expression and Adaptation in Bacteria is a comprehensive two-volume work bringing together both review and original research articles on key topics in stress and environmental control of gene expression in bacteria. Volume One contains key overview chapters, as well as content on one/two/three component regulatory systems and stress responses, sigma factors and stress responses, small non-coding RNAs and stress responses, toxin-antitoxin systems and stress responses, stringent response to stress, responses to UV irradiation, SOS and double stranded systems repair systems and stress, adaptation to both oxidative and osmotic stress, and desiccation tolerance and drought stress. Volume Two covers heat shock responses, chaperonins and stress, cold shock responses, adaptation to acid stress, nitrosative stress, and envelope stress, as well as iron homeostasis, metal resistance, quorum sensing, chemotaxis and biofilm formation, and viable but not culturable (VBNC) cells. Covering the full breadth of current stress and environmental control of gene expression studies and expanding it towards future advances in the field, these two volumes are a one-stop reference for (non) medical molecular geneticists interested in gene regulation under stress.