Author: Kakoli Bose Publisher: Springer Nature ISBN: 9811649871 Category : Medical Languages : en Pages : 315
Book Description
This book is immensely useful for graduate students as well as researchers to understand the basics of molecular biology and Recombinant DNA Technology. It provides a comprehensive overview of different approaches for the synthesis of recombinant proteins from E. coli including their cloning, expression and purification. Recent advances in genomics, proteomics, and bioinformatics have facilitated the use of Recombinant DNA Technology for evaluating the biophysical and biochemical properties of various proteins. The book starts with an introductory chapter on gene cloning, protein expression and purification and its implication in current research and commercial applications. Each chapter provides a lucid set of principles, tools and techniques for both students and instructors. The protocols described have been aptly exemplified, and troubleshooting techniques have been included to aid better understanding. Moreover, the set of questions at the end of each chapter have been particularly formulated to help effective learning.
Author: Ramnath Seetharam Publisher: CRC Press ISBN: 9780824782771 Category : Medical Languages : en Pages : 352
Book Description
Covering both new and traditional topics in the purification and analysis of recombinant proteins, this volume demonstrates how to overcome problems in protein research and presents practical methods used in protein work, explaining their theoretical bases. The collection also explores innovative co
Author: I.M. Rosenberg Publisher: Springer Science & Business Media ISBN: 1475711085 Category : Mathematics Languages : en Pages : 453
Book Description
This book is designed to be a practical progression of experimental techniques an investigator may follow when embarking on a biochemical project. The protocols may be performed in the order laid out or may be used inde pendently. The aim of the book is to assist a wide range of researchers. from the novice to the frustrated veteran, in the choice and design of experiments that are to be performed to provide answers to specific questions. The manual describes standard techniques that have been shown to work, as well as some newer ones that are beginning to prove important. By following the promi nently numbered steps. you can work your way through any protocol. whether it's a new technique or a task you've done before for which you need a quick review or updated methodology. This manual will assist the experimentalist in designing properly controlled experiments. There will be no advice for dealing with specific pieces of equip ment other than encouragement to read the manual, if you can find it. Through out all manipulations try to be objective. Be on the lookout for unexpected findings. You will learn the most from unexpected results. and they are often the beginning of the next project. It is never possible to record too much in your lab notebook. Do not get discouraged. Remember, things will not always run smoothly.
Author: Sharon A. Doyle Publisher: Humana Press ISBN: 9781617378218 Category : Science Languages : en Pages : 0
Book Description
Despite exciting advances in genome sequencing, isolating a protein from its expression system in its native form still presents a complex challenge. In High Throughput Protein Expression and Purification: Methods and Protocols, leading scientists detail the most successful protocols currently in use, including various high throughput cloning schemes, protein expression analysis, and production protocols. This volume describes the use of E. coli, insect, and mammalian cells, as well as cell-free systems for the production of a wide variety of proteins, including glycoproteins and membrane proteins, in order to best represent strategies that create and exploit common features to enable simplified cloning, stable expression, and purification of proteins. Written in the highly successful Methods in Molecular BiologyTM series format, the chapters present brief introductions to the subject, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and a Notes section for tips on troubleshooting and avoiding known pitfalls. Cutting-edge and comprehensive, High Throughput Protein Expression and Purification: Methods and Protocols is an ideal reference for protein biochemists and all those who wish to apply these easy-to-use protocols to the many applicable fields.
Author: Hafiz Ahmed Publisher: CRC Press ISBN: 0203507436 Category : Medical Languages : en Pages : 408
Book Description
Principles and Reactions of Protein Extraction, Purification, and Characterization provides the mechanisms and experimental procedures for classic to cutting-edge techniques used in protein extraction, purification, and characterization. The author presents the principles and reactions behind each procedure and uses tables to compare the different
Author: Allison R. Kermode Publisher: John Wiley & Sons ISBN: 1118801482 Category : Science Languages : en Pages : 496
Book Description
A single volume collection that surveys the exciting field of plant-made pharmaceuticals and industrial proteins This comprehensive book communicates the recent advances and exciting potential for the expanding area of plant biotechnology and is divided into six sections. The first three sections look at the current status of the field, and advances in plant platforms and strategies for improving yields, downstream processing, and controlling post-translational modifications of plant-made recombinant proteins. Section four reviews high-value industrial and pharmacological proteins that are successfully being produced in established and emerging plant platforms. The fifth section looks at regulatory challenges facing the expansion of the field. The final section turns its focus toward small molecule therapeutics, drug screening, plant specialized metabolites, and plants as model organisms to study human disease processes. Molecular Pharming: Applications, Challenges and Emerging Areas offers in-depth coverage of molecular biology of plant expression systems and manipulation of glycosylation processes in plants; plant platforms, subcellular targeting, recovery, and downstream processing; plant-derived protein pharmaceuticals and case studies; regulatory issues; and emerging areas. It is a valuable resource for researchers that are in the field of plant molecular pharming, as well as for those conducting basic research in gene expression, protein quality control, and other subjects relevant to molecular and cellular biology. Broad ranging coverage of a key area of plant biotechnology Describes efforts to produce pharmaceutical and industrial proteins in plants Provides reviews of recent advances and technology breakthroughs Assesses realities of regulatory and cost hurdles Forward looking with coverage of small molecule technologies and the use of plants as models of human disease processes Providing wide-ranging and unique coverage, Molecular Pharming: Applications, Challenges and Emerging Areas will be of great interest to the plant science, plant biotechnology, protein science, and pharmacological communities.
Author: Paul Cutler Publisher: Springer Science & Business Media ISBN: 159259655X Category : Science Languages : en Pages : 474
Book Description
The first edition of Protein Purification Protocols (1996), edited by Professor Shawn Doonan, rapidly became very successful. Professor Doonan achieved his aims of p- ducing a list of protocols that were invaluable to newcomers in protein purification and of significant benefit to established practitioners. Each chapter was written by an ex- rienced expert in the field. In the intervening time, a number of advances have w- ranted a second edition. However, in attempting to encompass the recent developments in several areas, the intention has been to expand on the original format, retaining the concepts that made the initial edition so successful. This is reflected in the structure of this second edition. I am indebted to Professor Doonan for his involvement in this new edition and the continuity that this brings. Each chapter that appeared in the original volume has been reviewed and updated to reflect advances and bring the topic into the 21st century. In many cases, this reflects new applications or new matrices available from vendors. Many of these have increased the performance and/or scope of the given method. Several new chapters have been introduced, including chapters on all the currently used protein fractionation and ch- matographic techniques. They introduce the theory and background for each method, providing lists of the equipment and reagents required for their successful execution, as well as a detailed description of how each is performed.
Author: Dermot Walls Publisher: Humana Press ISBN: 9781607619147 Category : Science Languages : en Pages : 527
Book Description
A prerequisite for elucidating the structure and function of any protein is the prior purification of that protein. This necessity has led to the development of many purification schemes and chromatographic methods for the isolation of native proteins from complex sources. In Protein Chromatography: Methods and Protocols, leading researchers present clear protocol-style chapters that are suitable for newcomers and experts alike. The book opens with vital topics in protein biochemistry, addressing such areas as protein stability and storage, avoiding proteolysis during chromatography, protein quantitation methods including immuno-qPCR, and the contrasting challenges that microfluidics and scale-up production pose to the investigator, and then it segues into key methods involving the generation and purification of recombinant proteins through recombinant antibody production and the tagging of proteins, amongst other means, as well as many variations on classic techniques such as ion-exchange and immunoaffinity chromatography. Written in the highly successful Methods in Molecular BiologyTM series format, protocols chapters include introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Protein Chromatography: Methods and Protocols will greatly aid scientists in establishing these essential techniques in their own laboratories and furthering our understanding of the many imperative functions of proteins.
Author: Eduardo A. Ceccarelli Publisher: Frontiers E-books ISBN: 2889192946 Category : Biotechnology Languages : en Pages : 103
Book Description
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.
Author: Kakoli Bose
Author: Ramnath Seetharam
Author: I.M. Rosenberg
Author: Sharon A. Doyle
Author: Hafiz Ahmed
Author: Renaud Vincentelli
Author: Allison R. Kermode
Author: Paul Cutler
Author: Dermot Walls
Author: Eduardo A. Ceccarelli