Insoluble Proteins: Methods and Protocols [Paperback]

This volume serves to aid researchers working in the recombinant protein production field by describing a wide number of protocols and examples. Chapters describe recombinant protein production in different expression systems, prokaryotic and eukaryotic expression systems, purification protocols, characterization of insoluble proteins and a general overview of interesting applications of insoluble proteins. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.

 Authoritative and cutting-edge, Insoluble Proteins: Methods and Protocols, Second Edition aims to be a useful practical guide to researches to help further their study in this field.

General Introduction: Recombinant protein production and purification of insoluble proteins.- Overcoming the solubility problem in E. coli: available approaches for recombinant protein production.- A3D 2.0 update for the prediction and optimization of protein solubility.- Tailoring codon usage to the underlying biology for protein expression optimization.- Protein Expression Optimization Strategies in E. coli: A Tailored Approach in Strain Selection and Parallelizing Expression Conditions.- A novel approach for production of aggregation prone proteins using the spidroin-derived NT* tag.- Cleavable Self-Aggregating Tags (cSAT) for Therapeutic Peptide Expression and Purification.- Use Intein Cleavable Polyhydroxyalkanoate Synthase Fusions to Improve Protein Solubility.- Introduction of a hexalysine (6K) tag can protect from N-terminal cleavage and increase yield of recombinant proteinslƒ