Recombinant Protein Expression: Prokaryotic hosts and cell-free systems

Recombinant Protein Expression: Prokaryotic hosts and cell-free systems

1st Edition - October 28, 2021
This is the Latest Edition
  • Editors: William O'Dell, Zvi Kelman
  • eBook ISBN: 9780323901475
  • Hardcover ISBN: 9780323901468

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Description

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.

Key Features

  • 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

Readership

Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists

Table of Contents

  • Cover image
  • Title page
  • Table of Contents
  • Copyright
  • Contributors
  • Preface
  • Bacterial hosts
  • Archaeal hosts
  • Cell-free systems
  • Section I: Bacterial hosts
  • Chapter One: Starting a new recombinant protein production project in Escherichia coli
  • Abstract
  • 1: Introduction
  • 2: Choosing an appropriate E. coli strain
  • 3: E. coli cultivation for recombinant protein production
  • 4: Methods
  • 5: Afterword
  • Acknowledgments
  • Chapter Two: From the notebook to recombinant protein production in Escherichia coli: Design of expression vectors and gene cloning
  • Abstract
  • 1: Introduction
  • 2: Tweaking the coding sequence of interest
  • 3: Selecting a suitable expression vector
  • 4: Working example: Design, construction, and cloning of a CDS
  • 5: Methods
  • 6: Afterword
  • Acknowledgments
  • Chapter Three: Use of tandem affinity–buffer exchange chromatography online with native mass spectrometry for optimizing overexpression and purification of recombinant proteins
  • Abstract
  • 1: Introduction
  • 2: Materials
  • 3: Methods
  • 4: Notes
  • 5: Discussion
  • 6: Summary
  • Chapter Four: Purification, reconstitution, and mass analysis of archaeal RNase P, a multisubunit ribonucleoprotein enzyme
  • Abstract
  • 1: Introduction
  • 2: Materials
  • 3: Methods
  • 4: Notes
  • 5: Conclusions
  • Acknowledgments
  • Chapter Five: Production of antibodies in SHuffle Escherichia coli strains
  • Abstract
  • 1: Introduction
  • 2: Equipment and materials
  • 3: Protocol
  • 4: Concluding remarks
  • 5: Notes
  • Chapter Six: Improved folding of recombinant protein via co-expression of exogenous chaperones
  • Abstract
  • 1: Introduction
  • 2: Methods
  • 3: Protocol for in vitro deconstruction assay
  • 4: Key resources
  • 5: Materials and equipment
  • 6: Step-by-step method details
  • 7: Morphological characterization of deconstruction assay products
  • 8: Structural characterization of deconstruction assay products
  • 9: Results
  • 10: Discussion
  • 11: Future development
  • Acknowledgments
  • Chapter Seven: Fusing an insoluble protein to GroEL apical domain enhances soluble expression in Escherichia coli
  • Abstract
  • 1: Introduction
  • 2: Before you begin
  • 3: Key resources table
  • 4: Materials and equipment
  • 5: Step-by-step method details
  • 6: Expected outcomes
  • 7: Advantages
  • 8: Limitations
  • Chapter Eight: Method for high-efficiency fed-batch cultures of recombinant Escherichia coli
  • Abstract
  • 1: Introduction
  • 2: Before you begin
  • 3: Materials and equipment
  • 4: Step-by-step method details
  • 5: Expected outcomes
  • 6: Advantages
  • 7: Limitations
  • 8: Safety considerations and standards
  • Acknowledgments
  • Chapter Nine: Fed-batch production of deuterated protein in Escherichia coli for neutron scattering experimentation
  • Abstract
  • 1: Introduction
  • 2: Materials and equipment
  • 3: Media preparation and cell adaption
  • 4: Fed-batch production of deuterated protein
  • 5: Expected outcomes
  • 6: Advantages
  • 7: Limitations
  • 8: Safety considerations and standards
  • 9: Alternative methods/procedures
  • Section II: Archaeal hosts
  • Chapter Ten: Thermococcus kodakarensis provides a versatile hyperthermophilic archaeal platform for protein expression
  • Abstract
  • 1: Introduction
  • 2: Genetic systems for Thermococcus kodakarensis
  • 3: Purifying proteins and identifying protein interactions
  • 4: Reagents, recipes and equipment
  • 5: Protocols
  • 6: Advantages and future perspectives
  • 7: Notes
  • Chapter Eleven: Recombinant protein expression in Sulfolobus islandicus
  • Abstract
  • 1: Introduction
  • 2: Equipment
  • 3: Materials
  • 4: Step-by-step protocol
  • 5: Expected outcomes
  • 6: Advantages
  • 7: Limitations
  • 8: Optimization and troubleshooting
  • Acknowledgments
  • Chapter Twelve: High-level synthesis and secretion of laccase, a metalloenzyme biocatalyst, by the halophilic archaeon Haloferax volcanii
  • Abstract
  • 1: Introduction
  • 2: Haloarchaea as a resource for biocatalysts
  • 3: Haloarchaea as platform for high-level protein secretion
  • 4: General strategy for Haloferax volcanii to secrete laccase
  • 5: Methods
  • 6: Protocol for laccase production using H. volcanii as the microbial platform
  • 7: Future perspectives
  • Chapter Thirteen: Expression and tandem affinity purification of 20S proteasomes and other multisubunit complexes in Haloferax volcanii
  • Abstract
  • 1: Introduction
  • 2: Proteasomes as multisubunit complexes
  • 3: General strategy for use of Haloferax volcanii to express and purify multisubunit complexes
  • 4: Protocols for heterologous expression and purification of multisubunit complexes
  • 5: Future perspectives
  • Acknowledgments
  • Chapter Fourteen: Purification and characterization of ribonucleoprotein effector complexes of Sulfolobus islandicus CRISPR-Cas systems
  • Abstract
  • 1: Introduction
  • 2: Before you begin
  • 3: Key resources table
  • 4: Materials and equipment
  • 5: Consumable items
  • 6: Step-by-step method details
  • 7: Expected outcomes
  • 8: Advantages
  • 9: Limitations
  • 10: Optimization and troubleshooting
  • 11: Safety considerations and standards
  • Acknowledgments
  • Section III: Cell-free systems
  • Chapter Fifteen: Guidelines for nucleic acid template design for optimal cell-free protein synthesis using an Escherichia coli reconstituted system or a lysate-based system
  • Abstract
  • 1: Introduction
  • 2: Requirements for protein synthesis from nucleic acid templates
  • 3: Types of templates
  • 4: Protocols
  • 5: Results
  • 6: Summary
  • Acknowledgment
  • Chapter Sixteen: Cell-free protein synthesis of CRISPR ribonucleoproteins (RNP)
  • Abstract
  • 1: Introduction
  • 2: DNA template design for Cas effector and guide RNA
  • 3: Materials and equipment
  • 4: Protocol
  • 5: Analysis
  • 6: Optimization and troubleshooting
  • 7: Conclusion
  • Acknowledgment
  • Chapter Seventeen: Leishmania tarentolae cell-free based approach for rapid anitbody–antigen interaction analysis
  • Abstract
  • 1: Introduction
  • 2: Materials
  • 3: Methods
  • 4: Summary
  • 5: Notes
  • 6: Protein sequences
  • Chapter Eighteen: Cell-free protein synthesis using Chinese hamster ovary cells
  • Abstract
  • 1: Introduction
  • 2: Key resources table
  • 3: Materials and equipment
  • 4: Step-by-step method details
  • 5: Expected outcomes
  • 6: Advantages
  • 7: Limitations
  • 8: Optimization and troubleshooting
  • 9: Safety considerations and standards
  • 10: Alternative methods/procedures

Product details

  • No. of pages: 460
  • Language: English
  • Copyright: © Academic Press 2021
  • Published: October 28, 2021
  • Imprint: Academic Press
  • eBook ISBN: 9780323901475
  • Hardcover ISBN: 9780323901468
  • About the Serial Volume Editors

    William O'Dell

    William Brad O'Dell is a PhD biologist experienced in in Molecular Biology, Protein Expression, Protein Purification, Crystallography, Lab-Scale Yeast Fermentation, Flow Cytometry and Small Angle Scattering of Molecules and Materials.

    Affiliations and Expertise

    Biologist, National Institute of Standards and Technology (NIST), MD, USA

    Zvi Kelman

    Zvi Kelman
    Zvi Kelman is the Director of the Biomolecular Labeling Laboratory (BL2), National Institute of Standards and Technology (NIST). He is also an Adjunct Professor in the Department of Cell Biology and Molecular Genetics at the University of Maryland, College Park and affiliated with the Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine. Zvi earned a B.Sc. in Agriculture from the Hebrew University in Jerusalem and a M.Sc. in Cell Biology from the Weizmann Institute of Science. After receiving a Ph.D. in Molecular Biology from Cornell University, he was a Helen Hay Whitney Foundation Post-Doctoral Fellow at Johns Hopkins University and Memorial Sloan-Kettering Cancer Center. Upon completion of his postgraduate training, he became a Life Technologies Professor at the Center for Advanced Research in Biotechnology (CARB), University of Maryland Biotechnology Institute (UMBI). Zvi moved to the University of Maryland, College Park in 2010 as a Professor in the Department of Cell Biology and Molecular Genetics. In 2011 he was recruited to NIST to direct the Biomolecular Labeling Laboratory.

    Affiliations and Expertise

    Director, Biomolecular Labeling Laboratory, NIST-IBBR, Rockville, MD, USA