Download or read book Textbook on Cloning Expression and Purification of Recombinant Proteins written by Kakoli Bose and published by Springer Nature. This book was released on 2022-01-25 with total page 315 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.

Download or read book Fundamentals of Recombinant Protein Production Purification and Characterization written by Deepti Yadav and published by Elsevier. This book was released on 2024-09-27 with total page 307 pages. Available in PDF, EPUB and Kindle. Book excerpt: Fundamentals of Recombinant Protein Production, Purification and Characterization is organized into nine chapters in a logical fashion that cover an introduction to recombinant proteins and expression in different host expression systems, extraction, purification and analysis of proteins. This important reference features protocols, along with the advantages and disadvantage of each expression hosts and characterization technique (presented in tabular format) and offers detailed coverage of all aspects of protein production and processing (upstream and downstream processing) in one place. Finally, the book ends with different characterization techniques. Production of recombinant proteins for biotechnological and therapeutic applications at a large scale is an essential need of mankind. With the huge application potential of therapeutic and industrial proteins, there has been increasing demand for effective and efficient bioprocessing strategies. Recent progress around recombinant DNA technologies and bioprocessing strategies has paved the way for efficient production of recombinant proteins. Important factors such as insolubility and cost of production need to be considered for large scale production of these recombinant proteins. Includes step-by-step reproducible protocols while also providing updated information on the rationale and latest developments in expression systems Can also be used as a handbook for protein expression and purification as expression systems and chromatographic methods are explained in detail Consists of notes on troubleshooting from the eminent researchers in the field Provides comprehensive information on protein production, purification and characterization in a single volume Describes different purification methods for comparatively difficult to obtain proteins Brings the topics of recombinant protein expression, purification and characterization together, thereby making it the first resource on how to solve problems with respect to upstream and downstream processing of heterologous proteins

Download or read book Guide to Protein Purification written by Richard R Burgess and published by Academic Press. This book was released on 2009-11-03 with total page 915 pages. Available in PDF, EPUB and Kindle. Book excerpt: Guide to Protein Purification, Second Edition provides a complete update to existing methods in the field, reflecting the enormous advances made in the last two decades. In particular, proteomics, mass spectrometry, and DNA technology have revolutionized the field since the first edition’s publication but through all of the advancements, the purification of proteins is still an indispensable first step in understanding their function. This volume examines the most reliable, robust methods for researchers in biochemistry, molecular and cell biology, genetics, pharmacology and biotechnology and sets a standard for best practices in the field. It relates how these traditional and new cutting-edge methods connect to the explosive advancements in the field. This "Guide to" gives imminently practical advice to avoid costly mistakes in choosing a method and brings in perspective from the premier researchers while presents a comprehensive overview of the field today. Gathers top global authors from industry, medicine, and research fields across a wide variety of disciplines, including biochemistry, genetics, oncology, pharmacology, dermatology and immunology Assembles chapters on both common and less common relevant techniques Provides robust methods as well as an analysis of the advancements in the field that, for an individual investigator, can be a demanding and time-consuming process

Download or read book Manipulation and Expression of Recombinant DNA written by Sue Carson and published by Elsevier. This book was released on 2005-12-15 with total page 172 pages. Available in PDF, EPUB and Kindle. Book excerpt: This manual is an indispensable tool for introducing advanced undergraduates and beginning graduate students to the techniques of recombinant DNA technology, or gene cloning and expression. The techniques used in basic research and biotechnology laboratories are covered in detail. Students gain hands-on experience from start to finish in subcloning a gene into an expression vector, through purification of the recombinant protein.The second edition has been completely re-written, with new laboratory exercises and all new illustrations and text, designed for a typical 15-week semester, rather than a 4-week intensive course. The “project approach to experiments was maintained: students still follow a cloning project through to completion, culminating in the purification of recombinant protein. It takes advantage of the enhanced green fluorescent protein—students can actually visualize positive clones following IPTG induction. *Cover basic concepts and techniques used in molecular biology research labs*Student-tested labs proven successful in a real classroom laboratories*Exercises simulate a cloning project that would be performed in a real research lab*"Project" approach to experiments gives students an overview of the entire process*Prep-list appendix contains necessary recipes and catalog numbers, providing staff with detailed instructions

Download or read book Recombinant Protein Expression Prokaryotic hosts and cell free systems written by and published by Academic Press. This book was released on 2021-10-29 with total page 462 pages. Available in PDF, EPUB and Kindle. Book excerpt: Recombinant Protein Expression, Part A, Volume 659 in the Methods in Enzymology series, highlights new advances in the field with this new volume presenting interesting chapters on Multiplexed analysis protein: Protein interactions of polypeptides translated in Leishmania cell-free system, MultiBac system and its applications, performance and recent, Production of antibodies in Shuffle, Designing hybrid-promoter architectures by engineering cis-acting DNA sites to enhance transcription in yeast, Designing hybrid-promoter architectures by engineering cis-acting DNA sites to deregulate transcription in yeast, Antibody or protein-based vaccine production in plants, Cell-free protein synthesis, Plant-based expression of biologic drugs, and much more. Additional sections cover the Use of native mass spectrometry to guide detergent-based rescue of non-native oligomerization by recombinant proteins, Advancing overexpression and purification of recombinant proteins by pilot optimization through tandem affinity-buffer exchange chromatography online with native mass spectrometry, Method for High-Efficiency Fed-batch cultures of recombinant Escherichia coli, Method to transfer Chinese hamster ovary (CHO) shake flask experiments to the ambr® 250, and Expression of recombinant antibodies in Leishmania tarentolae. Provides the authority and expertise of leading contributors from an international board of authors Presents the latest release in the Methods in Enzymology serial Updated release includes the latest information on Recombinant Protein Expression

Download or read book Recombinant Protein Expression Eukaryotic hosts written by and published by Academic Press. This book was released on 2021-11-04 with total page 384 pages. Available in PDF, EPUB and Kindle. Book excerpt: Recombinant Protein Expression, Part B, Volume 660 in the Methods in Enzymology series, highlights new advances in the field with this new volume presenting interesting chapters on Multiplexed analysis protein: Protein interactions of polypeptides translated in Leishmania cell-free system, MultiBac system and its applications, performance and recent, Production of antibodies in Shuffle, Designing hybrid-promoter architectures by engineering cis-acting DNA sites to enhance transcription in yeast, Designing hybrid-promoter architectures by engineering cis-acting DNA sites to deregulate transcription in yeast, Antibody or protein-based vaccine production in plants, Cell-free protein synthesis, Plant-based expression of biologic drugs, and much more. Additional sections cover the Use of native mass spectrometry to guide detergent-based rescue of non-native oligomerization by recombinant proteins, Advancing overexpression and purification of recombinant proteins by pilot optimization through tandem affinity-buffer exchange chromatography online with native mass spectrometry, Method for High-Efficiency Fed-batch cultures of recombinant Escherichia coli, Method to transfer Chinese hamster ovary (CHO) shake flask experiments to the ambr® 250, and Expression of recombinant antibodies in Leishmania tarentolae. Provides the authority and expertise of leading contributors from an international board of authors Presents the latest release in the Methods in Enzymology serial Updated release includes the latest information on Recombinant Protein Expression

Download or read book Recombinant protein expression in microbial systems written by Eduardo A. Ceccarelli and published by Frontiers E-books. This book was released on 2014-10-02 with total page 103 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.

Download or read book Cloning Gene Expression and Protein Purification written by Jennifer Edwards and published by Oxford University Press, USA. This book was released on 2001 with total page 435 pages. Available in PDF, EPUB and Kindle. Book excerpt: On the forefront of modern scientific innovation, Cloning, Gene Expression and Protein Purification: Experimental Procedures and Process Rationale effectively doubles as a laboratory manual for students and a reference book for professional researchers. Designed for advanced undergraduate and beginning graduate students in molecular biology, this unique combination lecture/laboratory resource presents detailed protocols for the multi-step process involved in isolating a gene, cloning and characterizing it, expressing its encoded protein, and purifying and characterizing the protein's basic physical properties. This manageable volume includes both theoretical background and practical procedures and is structured around twenty experiments that demonstrate how to prepare, manipulate, and analyze plasmids, produce fusion proteins in bacteria, and purify these proteins based on unique chemical properties or substrate affinities. The book describes advanced topics such as the use of antibodies and the techniques developed to transform their structures, as well as combinatorial approaches designed to manipulate the structure and functions of proteins and nucleic acids. Supplemental literature provides a variety of theoretical explanations encouraging a more intuitive understanding of the experimental mechanisms and behaviors of the chemical participants, while also giving students the tools needed to become "capable proactive researchers." Features: � Emphasizes electrophoresis, Southern and Western blotting, and combinatorial techniques � Defines clear reaction mechanisms; stipulates the functions of reagents; and helps students think about the precise consequences of solution and procedural manipulations � Discusses fluorophores, and solvent effects on protein structure � Characterizes plasmids, cDNAs, and antibody probes (available from ATCC) in research literature � Includes carefully selected primary source research literature and articles from current vendor literature � Contains a glossary of unfamiliar phrases and jargon; important summary statements and conclusions are italicized � Provides an alphabetized list of common reagents for rapid reference � Offers an extensive index of concepts and terms � Categorizes helpful and distinctive information into five types of supplemental literature: Innovation/ Insight, Theory/Principle, Process Rationale, Vendor Literature, and Alternative Approaches

Download or read book Gene Cloning Expression and Purification of Membrane Proteins written by Paola Luci and published by . This book was released on 2003 with total page 111 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Gene Cloning and Expression Technologies written by Michael P. Weiner and published by Biotechniques Press. This book was released on 2002 with total page 652 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Recombinant Protein Expression in Mammalian Cells written by David L. Hacker and published by Springer Nature. This book was released on with total page 359 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Purification and Analysis of Recombinant Proteins written by Ramnath Seetharam and published by CRC Press. This book was released on 1991-01-07 with total page 352 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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

Download or read book Recombinant Proteins in Plants written by Stefan Schillberg and published by . This book was released on 2022 with total page 346 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume provided methods and protocols on recombinant protein production in different plant systems, downstream processing, and strategies to optimize protein expression. Chapters guide readers through recombinant protein production in important plant systems, protein recovery and purification, different strategies to optimise productivity, cloning and fusion protein approaches, and the regulation and freedom to operate analysis of plant-produced proteins. 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, Recombinant Proteins in Plants: Methods and Protocols aims to be useful to newcomers and experienced researchers interested in expanding their expertise in the field of plant-based protein production. Chapters 6, 8 and 17 are available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

Download or read book Production of Recombinant Proteins written by Gerd Gellissen and published by John Wiley & Sons. This book was released on 2006-03-06 with total page 429 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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."

Download or read book Molecular Cloning Expression Purification and Immunological Characterization of Proteins Encoded by Regions of Difference Genes of Mycobacterium Tuberculosis written by Shumaila Nida Muhammad Hanif and published by . This book was released on 2011 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Production Technology of Recombinant Therapeutic Proteins written by Chiranjib Chakraborty and published by Daya Books. This book was released on 2004 with total page 290 pages. Available in PDF, EPUB and Kindle. Book excerpt: An Increasing Number Of Recombinant Therapeutic Proteins Are Currently Being Developed, Tested In Clinical Trials And Marketed For Used. Most Of The Recombinant Therapeutic Proteins Are Being Successfully Produced Into Escherichia Coli And Pichia Pastoris Expression System. These Two Expression Systems Are Very Much Efficient And Cost Effective. This Book Takes A Close Look Of These Two Expression Systems And Fermentation Conditions, Purification Strategies Of Different Recombinant Proteins. This Book Also Discusses The Market Size And Cost Analysis For The Production Of Different Therapeutic Proteins And Some General Experimental Protocols For Production. Contents Part I: Recombinant Protein Expression Into Escherichia Coli And Fermentation Conditions; Chapter 1: Introduction; Chapter 2: Construction Of Efficient Expression Vector (Plasmid); Chapter 3: Factors Affecting Transcription, Promoters, Upstream Elements, Transcriptional Terminators, Transcriptional Antitermin, Tightly Regulated Expression Systems; Chapter 4: Mrna Stability; Chapter 5: Factors Affecting Translation, Mrna Translational Initiator, Translational Enhancers, Translational Termination; Chapter 6: Expression Of Target Protein And The Compartments Of Expression, Cytoplasmic Expression, Periplasmic Expression, Extracellular Secretion; Chapter 7: Fusion Proteins; Chapter 8: Post-Translational Protein Folding; Chapter 8: Codon Usage; Chapter 10: Protein Degradation; Chapter 11: Fermentation Conditions For High-Density Cell Cultivation (Hdcc), Growth Medium, Efficient Production Of Recombinant Protein In Hdcc, Nutrient Feeding Strategy In Hdcc; Chapter 12: One Examples Of Protein Production Using E. Coli Expression System; Chapter 13: Conclusion. Part Ii: Recombinant Protein Expression Into Yeast, Pichia Pastoris And Fermentation Conditions; Chapter 1: Introduction; Chapter 2: Why P. Pastoris? Chapter 3: Construction Of Expression Strains, Expression Vectors, Alternative Promoters, Host Strains, Methanol Utilisation Phenotype, Protease-Reduced Host Strains, Integration Of Expression Vectors Into The P. Pastoris Genome, Generating Multicopy Strains; Chapter 4: Post-Translational Modifications Of Secreted Proteins, Secretion Signal Selection, N-Linked Glycosylation; Chapter 5: Production Of Recombinant Proteins In Fermenter Cultures Of The Yeast, Pichia Pastoris, Conceptual Basis For The P. Pastoris Expression System, High-Level Expression In Fermenter Cultures, Protein-Specific Adjustments To Improve Yield, Glycosylation Of Recombinant Proteins, Secretion Signals; Chapter 6: One Examples Of Protein Producing Using P. Pastoris Expression System, Chapter 7: Conclusion. Part Iii: Purification Strategies For Recombinant Proteins; Chapter 1: Purification Of Proteins; Chapter 2: Conventional Chromatography, Ion Exchange Chromatography, Reversed Phase Chromatography, Gel Permeation Chromatography, Affinity Chromatography, Affinity Tags, Cleavage, Conclusion. Part Iv: Market Size And Cost Analysis For The Production Of Therapeutic Proteins; Chapter 1: Market Size Of Therapeutic Proteins; Chapter 2: Outline Structure Of A Productin Unit And Cost Analysis For The Production Of Three Therapeutic Proteins. Part V: General Experimental Protocols; Chapter 1: Different Experimental Protocols, Preparation Of Genome Dna For E. Coli, A Differnt Method For Preparation Of Genomic Dna From Bacteria, Preparation Of Proteins From Periplasm (Osmotic Shock Method), Preparation Of Proteins From Outer Membrane, Transformation Of Plasmid Dna Into E. Coli (Calcium Chloride/Heat Shock Method), Transformation Of Plasmid Dna Into E. Coli (Electroporation), Sds-Page For Large Proteins, Sds-Page For Small Peptide, Pcr Amplification Of Dna, Protein Quantification: Brandford Method, Trans-Bloting For Protein, Restriction Enzyme Digestion Of Dna, Phenol/Chloroform Extraction Of Dna, Ethanol Precipitation Of Dna, Agarose Gel Electrophoresis, Transformation Of E. Coli By Electroporation (Alternative Method), Wizard Tm Pcr Preps Dna Purification System For Rapid, Purification Of Dna Fragments, Alternate Method For Purifying Dna From Agarose Gels, Southern Blotting, Rt Pcr Protocol, Using Superscript Reverse Transcriptase, Preparation Of Sequencing Gels, Isolation Of Rna From Mammalian Cells Using Rnazoltm (Teltest), Preparation For Yeast Transformation, Yeast Transformation, Digesting Prsq-Ura3 With Bamhi, Genomic Dna Preparation Of Yeast, Ligation (Circularisation) Of Genomic Dna Fragments, E. Coli Transformation (Alternate Method), Dna Miniprep From E. Coli (Alternate Method), Basic Plasmid Dna Isolation Protocol, Identification And Determination Of Amount Rec-Hum Proteins Via An Immunoenzymatic Test (Elisa), Determination Of Host Dna Contaminant Into R Hu Protein Through Dot Blot Method, Protocols For Down-Stream Processing.

Download or read book Understanding DNA and Gene Cloning written by Karl Drlica and published by John Wiley & Sons. This book was released on 2004 with total page 388 pages. Available in PDF, EPUB and Kindle. Book excerpt: Publisher's description: With DNA and gene cloning all over the news, readers need to understand the ongoing genetic revolution. In this highly acclaimed guide, Karl Drlica fully explains the basic science and technology readers need to understand the issues and make crucial decisions. Each step of the way he explains complex topics using easy-to-understand analogies.