Heterologous Gene Expression in E.Coli: Methods and Protocols

ISBN-13: 9781617379666 / Angielski / Twarda / 2010 / 310 str.

JR. Thomas C. Evans; Ming-Qun Xu

Opis

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

Recenzje

From the reviews:

"The editors bring together a diverse set of methods and examples demonstrating that, in fact, there is still plenty of active and exiting research pushing back some of the aforementioned limitations of E. coli. ... If recombinant protein expression is an important part of your research programme, either in an academic or industrial setting, then this book is worth a read - it could save you a fortune!" (Gavin Thomas, Microbiology Today, November, 2011)

Spis treści

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 Protein–Protein 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

Blurb

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.