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Heterologous Gene Expression in E coli Methods and Protocols 1st Edition by Nicola Burgess Brown 9781493968879 1493968874

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Heterologous Gene Expression in E coli Methods and Protocols 1st Edition Nicola A. Burgess-Brown (Eds.) Digital Instant Download

Author(s): Nicola A. Burgess-Brown (eds.)
ISBN(s): 9781493968879, 1493968874
Edition: 1
File Details: PDF, 11.18 MB
Year: 2017
Language: english
SKU: EB-5880984 Category: Tag:

Heterologous Gene Expression in E coli Methods and Protocols 1st Edition by Nicola Burgess Brown – Ebook PDF Instant Download/Delivery: 9781493968879, 1493968874

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Product details: 

ISBN 10: 1493968874

ISBN 13: 9781493968879

Author: 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:

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

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

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

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

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

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

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

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

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

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

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

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

  13. Protein Folding Using a Vortex Fluidic Device

  14. Removal of Affinity Tags with TEV Protease

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

  16. Production of Protein Kinases in E. coli

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

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

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

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

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

  22. In Vivo Biotinylation of Antigens in E. coli

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

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

  25. Mass Spectrometric Analysis of Proteins

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

  27. Interfacing Biocompatible Reactions with Engineered Escherichia coli

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Tags: Nicola Burgess Brown, Heterologous, Methods