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Heterologous Gene Expression in E.coli

  • Kartonierter Einband
  • 324 Seiten
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Practical and cutting-edge, Heterologous Gene Expression in E. coli: Methods and Protocols seeks to familiarize the researcher wit... Weiterlesen
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Beschreibung

Practical and cutting-edge, Heterologous Gene Expression in E. coli: Methods and Protocols seeks to familiarize the researcher with the myriad of E. coli expression strains available and move E. coli closer to that ideal of the perfect host.

Protein expression in a heterologous host is a cornerstone of biomedical research and of the biotechnology industry. Despite the advanced state of protein expression technology improvements are still needed. For example, membrane proteins constitute a significant percentage of the total cellular proteins but as a class are very difficult to overexpress, especially in a heterologous host. The ideal host would have the ability to express any protein, with relevant post-translational modifications, and be as easy to work with as E. coli. In Heterologous Gene Expression in E. coli: Methods and Protocols, expert scientists intimately familiar with the relevant techniques offer chapters that greatly expand the utility of this expression host. The contributions in this detailed volume describe methods, for example, to successfully express proteins in E. coli that would otherwise form aggregates in this host, to add post-translational modifications, to incorporate non-standard amino acid residues or moieties into E. coli expressed proteins, to identify binding partners, and to express membrane proteins. Written in the highly successful Methods in Molecular Biology™ format, chapters include introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Practical and cutting-edge, Heterologous Gene Expression in E. coli: Methods and Protocols seeks to familiarize the researcher with the myriad of E. coli expression strains available and move E. coli closer to that ideal of the perfect host.

Helps to guide scientists toward an ideal expression host

Provides easy to follow protocols, ready to be implemented in the lab

Includes tips from the experienced, expert contributors on troubleshooting common methodological pitfalls



Inhalt
1. Adjustment of Codon Usage Frequencies by Codon Harmonization Improves Protein Expression and Folding Evelina Angov, Patricia M. Legler, and Ryan M. Mease 2. SUMO Fusion Technology for Enhanced Protein Expression and Purification in Prokaryotes and Eukaryotes Yilin Yan, James Strickler, and Tauseef Butt 3. Molecular and Chemical Chaperones for Improving the Yields of Soluble Recombinant Proteins Ario de Marco 4. Genetic Selection of Solubility-Enhanced Proteins Using the Twin-Arginine Translocation System Adam C. Fisher, Mark A. Rocco, and Matthew P. DeLisa 5. Protein Folding Liquid Chromatography Quan Bai and Xindu Geng 6. Site-Specific Protein Labeling by Intein-Mediated Protein Ligation Inca Ghosh, Nancy Considine, Elissa Maunus, Luo Sun, Aihua Zhang, John Buswell, Thomas C. Evans, Jr., and Ming-Qun Xu 7. Efficient Expression of Human Aromatase (CYP19) in E. coli Norio Kagawa 8. Expression of Recombinant Cytochromes c in E. coli Yuri Y. Londer 9. Semi-Synthesis of Glycoproteins from E. coli through Native Chemical Ligation Jonathan P. Richardson and Derek Macmillan 10. Expression of Recombinant Proteins with Uniform N-Termini Orsolya Király, Lan Guan, and Miklós Sahin-Tóth 11. Recent Developments in Difficult Protein Expression: A Guide to E. coli Strains, Promoters, and Relevant Host Mutations James C. Samuelson 12. Periplasmic Chaperones Used to Enhance Functional Secretion of Proteins in E. coli Martin Schlapschy and Arne Skerra 13. Engineering Unusual Amino Acids into Peptides Using Lantibiotic Synthetase Jun-ichi Nagao, Kouki Shioya, Yoshitaka Harada, Ken-ichi Okuda, Takeshi Zendo, Jiro Nakayama, and Kenji Sonomoto 14. The Targeted Expression of Nucleotide Sugar Transporters to the E. coli Inner Membrane Joe Tiralongo and Andrea Maggioni 15. Detection of ProteinProtein Interactions in Bacteria by GFP-Fragment Reconstitution Akira Kanno, Takeaki Ozawa, and Yoshio Umezawa 16. Enhancing the Solubility of Recombinant Proteins in Escherichia coli by Using Hexahistidine-Tagged Maltose-Binding Protein as a Fusion Partner Ping Sun, Joseph E. Tropea, and David S. Waugh 17. Introducing Predetermined Mutations throughout a Target Gene Using TDEM (Transposon Directed Base-Exchange Mutagenesis) Yun Cheol Kim 18. Fluorescent Site-Specific Labeling of Escherichia coli Expressed Proteins with Sfp Phosphopantetheinyl Transferase Aihua Zhang, Luo Sun, John Buswell, Nancy Considine, Inca Ghosh, Anastasiya Masharina, Christopher Noren, and Ming-Qun Xu

Produktinformationen

Titel: Heterologous Gene Expression in E.coli
Untertitel: Methods and Protocols
Editor:
EAN: 9781493957255
ISBN: 1493957252
Format: Kartonierter Einband
Herausgeber: Humana Press
Genre: Medizin
Anzahl Seiten: 324
Gewicht: 612g
Größe: H254mm x B178mm x T17mm
Jahr: 2016
Auflage: Softcover reprint of the original 1st ed. 2011

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