Heterologous Gene Expression in E.coli : Methods and Protocols / edited by Thomas C. Evans, Jr., Ming-Qun Xu.

Format:

Book

Contributor:

Evans, Jr., Thomas C., editor.

Xu, Ming-Qun, editor.

SpringerLink (Online service)

Series:

Methods in Molecular Biology, Methods and Protocols, 1064-3745 ; 705.

Springer Protocols (Springer-12345)

Methods in Molecular Biology, Methods and Protocols, 1064-3745 ; 705

Language:

English

Subjects (All):

Medicine.

Human genetics.

Proteomics.

Biomedicine.

Human Genetics.

Local Subjects:

Biomedicine.

Human Genetics.

Proteomics.

Physical Description:

1 online resource (XI, 310 pages).

Contained In:

Springer eBooks

Place of Publication:

Totowa, NJ : Humana Press, 2011.

System Details:

text file PDF

Summary:

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.

Contents:

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.

Other Format:

Printed edition:

ISBN:

9781617379673

Access Restriction:

Restricted for use by site license.