Insoluble Proteins

Methods and Protocols

Häftad, Engelska, 2022

Av Elena Garcia Fruitós, Anna Arís Giralt, Elena Garcia Fruitos, Anna Aris Giralt

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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.

Produktinformation

Utgivningsdatum2022-01-29
Mått178 x 254 x 30 mm
Vikt1 025 g
FormatHäftad
SpråkEngelska
SerieMethods in Molecular Biology
Antal sidor533
Upplaga2
FörlagSpringer-Verlag New York Inc.
ISBN9781071618615

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Biovetenskap inom Naturvetenskap och teknik

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 proteins expressed in the periplasm of E. coli.- Two-tiered selection and screening strategy to increase functional enzyme production in Escherichia coli.- Method for production of cysteine-rich proteins in Lactococcus lactis expression system.- Heterologous protein production in Lactobacillus (plantarum) using pSIP vectors.- Soluble recombinant protein production in Pseudoaltermonas haloplanktis TAC125: the case study of the full-length human CDKL5 protein.- A protocol to enhance soluble protein expression in the cytoplasm of Bacillus subtilis.- Applications of cell-free synthesized membrane protein precipitates.- High cell-density expression system: yeast cells in a phalanx efficiently produce a certain range of “difficult-to-express” secretory recombinant proteins.- Insect Cells-Baculovirus system for the production of difficult to express proteins: from expression screening for soluble constructs to protein quality control.- Transient gene expression in human Expi293 cells.- Recombinant Expression of Complex Proteins in Human Cells Lines.- Optimizing chaperone removal strategy from over-expressed recombinant proteins: GNE, a case study.- Intrinsically disordered proteins (IDP): purification under denaturing conditions.- Solubilization and Refolding of Inclusion Body Proteins.- Non-denaturing solubilization of Inclusion Bodies from Lactic Acid Bacteria.- Purification of Inclusion Bodies Produced in Bacteria and Yeast.- Eukaryotic aggresomes: protocols and tips for their production, purification and handling.- Characterization of the conformational properties of soluble and insoluble proteins by Fourier transform infrared spectroscopy.- Characterizing Soluble Protein Aggregates using Native Mass Spectrometry Coupled with Temperature-control Electrospray Ionization and Size-exclusion Chromatography.- Quality control of proteins solubilized from inclusion bodies.- Methods for the Characterizationof Protein Aggregates.- Insoluble Protein Applications: The Use of Bacterial Inclusion Bodies as Biocatalysts.- Methods for Processing Protein Aggregates into Surfaces.