Heterologous Gene Expression in E.coli
Methods and Protocols
(Sprache: Englisch)
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...
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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(TM) 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.
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.
Klappentext zu „Heterologous Gene Expression in E.coli “
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(TM) 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.
Inhaltsverzeichnis zu „Heterologous Gene Expression in E.coli “
Adjustment of Codon Usage Frequencies by Codon Harmonization Improves Protein Expression and Folding.- SUMO Fusion Technology for Enhanced Protein Expression and Purification in Prokaryotes and Eukaryotes.- Molecular and Chemical Chaperones for Improving the Yields of Soluble Recombinant Proteins.- Genetic Selection of Solubility-Enhanced Proteins Using the Twin-Arginine Translocation System.- Protein Folding Liquid Chromatography.- Site-Specific Protein Labeling by Intein-Mediated Protein Ligation.- Efficient Expression of Human Aromatase (CYP19) in E. coli.- Expression of Recombinant Cytochromes c in E. coli.- Semi-Synthesis of Glycoproteins from E. coli through Native Chemical Ligation.- Expression of Recombinant Proteins with Uniform N-Termini.- Recent Developments in Difficult Protein Expression: A Guide to E. coli Strains, Promoters, and Relevant Host Mutations.- Periplasmic Chaperones Used to Enhance Functional Secretion of Proteins in E. coli.- Engineering Unusual Amino Acids into Peptides Using Lantibiotic Synthetase.- The Targeted Expression of Nucleotide Sugar Transporters to the E. coli Inner Membrane.- Detection of Protein-Protein Interactions in Bacteria by GFP-Fragment Reconstitution.- Enhancing the Solubility of Recombinant Proteins in Escherichia coli by Using Hexahistidine-Tagged Maltose-Binding Protein as a Fusion Partner.- Introducing Predetermined Mutations throughout a Target Gene Using TDEM (Transposon Directed Base-Exchange Mutagenesis).- Fluorescent Site-Specific Labeling of Escherichia coli Expressed Proteins with Sfp Phosphopantetheinyl Transferase.
Bibliographische Angaben
- 2010, XI, 310 Seiten, Masse: 18,3 x 26 cm, Gebunden, Englisch
- Herausgegeben: Thomas C. Evans, Ming-Qun Xu
- Verlag: Springer, Berlin
- ISBN-10: 1617379662
- ISBN-13: 9781617379666
- Erscheinungsdatum: 02.12.2010
Sprache:
Englisch
Rezension zu „Heterologous Gene Expression in E.coli “
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)
Pressezitat
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)
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