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Textbook on Cloning, Expression and Purification of Recombinant Proteins

Textbook on Cloning, Expression and Purification of Recombinant Proteins
Author: Kakoli Bose
Publisher: Springer Nature
Total Pages: 315
Release: 2022-01-25
Genre: Medical
ISBN: 9811649871

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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.


Manipulation and Expression of Recombinant DNA

Manipulation and Expression of Recombinant DNA
Author: Sue Carson
Publisher: Elsevier
Total Pages: 172
Release: 2005-12-15
Genre: Science
ISBN: 0080456545

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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


Recombinant Protein Expression: Prokaryotic hosts and cell-free systems

Recombinant Protein Expression: Prokaryotic hosts and cell-free systems
Author:
Publisher: Academic Press
Total Pages: 462
Release: 2021-10-29
Genre: Science
ISBN: 0323901476

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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


Guide to Protein Purification

Guide to Protein Purification
Author: Richard R Burgess
Publisher: Academic Press
Total Pages: 915
Release: 2009-11-03
Genre: Science
ISBN: 0080923178

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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


Cloning, Gene Expression, and Protein Purification

Cloning, Gene Expression, and Protein Purification
Author: Jennifer Edwards
Publisher: Oxford University Press, USA
Total Pages: 435
Release: 2001
Genre: Science
ISBN: 9780195132946

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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


Recombinant Proteins in Plants

Recombinant Proteins in Plants
Author: Stefan Schillberg
Publisher:
Total Pages: 346
Release: 2022
Genre: Botany
ISBN: 9781071622414

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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.


High Throughput Protein Expression and Purification

High Throughput Protein Expression and Purification
Author: Sharon A. Doyle
Publisher: Humana Press
Total Pages: 0
Release: 2010-11-19
Genre: Science
ISBN: 9781617378218

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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.


Recombinant Protein Expression in Mammalian Cells

Recombinant Protein Expression in Mammalian Cells
Author: David L. Hacker
Publisher:
Total Pages: 311
Release: 2018
Genre: Biochemistry
ISBN: 9781493987306

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This detailed volume explores advances in vector design, DNA delivery, cell cultivation, host cell engineering, and bioprocess optimization within the study of recombinant protein expression in mammalian cells. The majority of the protocols employ either Chinese hamster ovary cells (CHO) or human embryonic kidney 293 cells (HEK293), the workhorses of the field, as the production host; however, the methods can be adapted to other mammalian hosts under the appropriate cell-specific conditions. 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 convenient, Recombinant Protein Expression in Mammalian Cells: Methods and Protocols aims to aid researchers in building on our knowledge of protein structure and function and to speed the discovery of new therapeutic proteins. Chapter 19 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.