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Heterologous Gene Expression in E.coli Nicola A. Burgess-Brown

Heterologous Gene Expression in E.coli By Nicola A. Burgess-Brown

Heterologous Gene Expression in E.coli by Nicola A. Burgess-Brown


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Summary

This detailed volume provides a toolbox for designing constructs, tackling expression and solubility issues, handling membrane proteins and protein complexes, and exploring innovative engineering of E.

Heterologous Gene Expression in E.coli Summary

Heterologous Gene Expression in E.coli: Methods and Protocols by Nicola A. Burgess-Brown

This detailed volume provides a toolbox for designing constructs, tackling expression and solubility issues, handling membrane proteins and protein complexes, and exploring innovative engineering of E. coli. The topics are largely grouped under four parts: high-throughput cloning, expression screening, and optimization of expression conditions, protein production and solubility enhancement, case studies to produce challenging proteins and specific protein families, as well as applications of E. coli expression. 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, Heterologous Gene Expression in E. coli: Methods and Protocols serves molecular biologists, biochemists and structural biologists, those in the beginning of their research careers to those in their prime, to give both an historical and modern overview of the methods available to express their genes of interest in this exceptional organism.

Table of Contents

Part I: High-Throughput Cloning, Expression Screening, and Optimization

1. Recombinant Protein Expression in E. coli: A Historical Perspective

Opher Gileadi

2. N- and C-Terminal Truncations to Enhance Protein Solubility and Crystallization: Predicting Protein Domain Boundaries with Bioinformatics Tools

Christopher D.O. Cooper and Brian D. Marsden

3. Harnessing the Profinity eXactTM System for Expression and Purification of Heterologous Proteins in E. coli

Yoav Peleg, Vadivel Prabahar, Dominika Bednarczyk, and Tamar Unger

4. ESPRIT: A Method for Defining Soluble Expression Constructs in Poorly Understood Gene Sequences

Philippe J. Mas and Darren J. Hart

5. Optimizing Expression and Solubility of Proteins in E. coli Using Modified Media and Induction Parameters

Troy Taylor, John-Paul Denson, and Dominic Esposito

6. Optimization of Membrane Protein Production Using Titratable Strains of E. coli

Rosa Morra, Kate Young, David Casas-Mao, Neil Dixon, and Louise E. Bird

7. Optimizing E. coli-Based Membrane Protein Production Using Lemo21(DE3) or pReX and GFP-Fusions

Grietje Kuipers, Markus Peschke, Nurzian Bernsel Ismail, Anna Hjelm, Susan Schlegel, David Vikstroem, Joen Luirink, and Jan-Willem de Gier

8. High Yield of Recombinant Protein in Shaken E. coli Cultures with Enzymatic Glucose Release Medium EnPresso B

Kaisa Ukkonen, Antje Neubauer, Vinit J. Pereira, and Antti Vasala

Part II: Protein Purification and Solubility Enhancement

9. A Generic Protocol for Purifying Disulphide-Bonded Domains and Random Protein Fragments Using Fusion Proteins with SUMO3 and Cleavage by SenP2 Protease

Huseyin Besir

10. A Strategy for Production of Correctly Folded Disulfide-Rich Peptides in the Periplasm of E. coli

Natalie J. Saez, Ben Cristofori-Armstrong, Raveendra Anangi, and Glenn F. King

11. Split GFP Complementation as Reporter of Membrane Protein Expression and Stability in E. coli: A Tool to Engineer Stability in a LAT Transporter

Ekaitz Errasti-Murugarren, Arturo Rodriguez-Banqueri, and Jose Luis Vazquez-Ibar

12. Acting on Folding Effectors to Improve Recombinant Protein Yields and Functional Quality

Ario de Marco

13. Protein Folding Using a Vortex Fluidic Device

Joshua Britton, Joshua N. Smith, Colin L. Raston, and Gregory A. Weiss

14. Removal of Affinity Tags with TEV Protease

Sreejith Raran-Kurussi, Scott Cherry, Di Zhang, and David S. Waugh

Part III: Case Studies to Produce Challenging Proteins and Specific Protein Families

15. Generation of Recombinant N-Linked Glycoproteins in E. coli

Benjamin Strutton, Stephen R.P. Jaffe, Jagroop Pandhal, and Phillip C. Wright

16. Production of Protein Kinases in E. coli

Charlotte A. Dodson

17. Expression of Prokaryotic Integral Membrane Proteins in E. coli

James D. Love

18. Multiprotein Complex Production in E. coli: The SecYEG-SecDFYajC-YidC Holotranslocon

Imre Berger, Quiyang Jiang, Ryan J. Schulze, Ian Collinson, and Christiane Schaffitzel

19. Membrane Protein Production in E. coli Lysates in Presence of Pre-Assembled Nanodiscs

Ralf-Bernhardt Rues, Alexander Grawe, Erik Henrich, and Frank Bernhard

20. Not Limited to E. coli: Versatile Expression Vectors for Mammalian Protein Expression

Katharina Karste, Maren Bleckmann, and Joop van den Heuvel

21. A Generic Protocol for Intracellular Expression of Recombinant Proteins in Bacillus subtilis

Trang Phan, Huynh Phuong, Tuom Truong, and Hoang Nguyen

Part IV: Applications of E. coli Expression

22. In Vivo Biotinylation of Antigens in E. coli

Susanne Graslund, Pavel Savitsky, and Susanne Muller-Knapp

23. Cold-Shock Expression System in E. coli for Protein NMR Studies

Toshihiko Sugiki, Toshimichi Fujiwara, and Chojiro Kojima

24. High-Throughput Production of Proteins in E. coli for Structural Studies

Charikleia Black, John J. Barker, Richard B. Hitchman, Hok Sau Kwong, Sam Festenstein, and Thomas B. Acton

25. Mass Spectrometric Analysis of Proteins

Rod Chalk

26. How to Determine Interdependencies of Glucose and Lactose Uptake Rates for Heterologous Protein Production with E. coli

David J. Wurm, Christoph Herwig, and Oliver Spadiut

27. Interfacing Biocompatible Reactions with Engineered Escherichia coli

Stephen Wallace and Emily P. Balskus

Additional information

NLS9781493983285
9781493983285
1493983288
Heterologous Gene Expression in E.coli: Methods and Protocols by Nicola A. Burgess-Brown
New
Paperback
Humana Press Inc.
2018-07-20
429
N/A
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