Showing papers in "Protein Expression and Purification in 2009"

TL;DR: A novel protein tag engineered to enhance expression and solubility of recombinant proteins and to provide efficient protein purification coupled with tag removal, designed to bind rapidly and covalently with a unique synthetic linker to achieve an essentially irreversible attachment.

TL;DR: Recombinant viscotoxins prepared using this protocol are toxic to HeLa cells and preserve the activity differences between isoforms B and A3 found in native proteins.

TL;DR: A new system which combines improved expression, solubility screening and purification efficiency, based on two newly constructed vectors, pEHISTEV and pEHISGFPTEV derived from a pET vector, which has the potential to be broadly applicable for structural biology.

TL;DR: A fungal strain, BCC2871 (Periconia sp.), was found to produce a thermotolerant beta-glucosidase, BGL I, with high potential for application in biomass conversion and Thermostability of the enzyme was improved remarkably in the presence of cellobiose, glucose, or sucrose.

TL;DR: A major breakthrough in transient gene expression (TGE) technologies was achieved by switching to other host cell lines, such as HEK293 and CHO lines, and the development of large scale transfection protocols using calcium-phosphate and polyethylenimine (PEI) as a carrier of plasmid DNA.

TL;DR: The TIPS method eliminates the need and protracted time for titering virus supernatants, and provides stable, concentrated storage of recombinant baculovirus in the form of infected cells, which is particularly advantageous for virus stocks which are unstable, such as those for sGC, and providing a highly efficient alternative for bacULovirus storage and expression.

TL;DR: In this paper, transient expression, purification and characterization of six chimeric heavy chain antibodies (cHCAbs) or fusion of camelid single domain antibodies (sdAbs) to human fragment crystallizable (Fc) are presented.

TL;DR: The strategy employed and the utility of thermostability assays are described in assessing how point mutations, truncations, detergents and ligands combine to develop a construct that forms diffraction-grade crystals.

TL;DR: The polypeptide analysis of the purified R-phycoerythrin by SDS-PAGE demonstrated that it contains four chromophore-carrying subunits and no colorless polypePTide and has two hexameric aggregates.

TL;DR: The results demonstrate the use of a cost effective prokaryotic expression system for generation and in vitro characterization of plant cysteine-rich proteins with potential antimicrobial activities against a wide range of phytopathogenic microorganisms in order to select the most effective agents for future in vivo studies.

TL;DR: By using a commercially available Q XL resin in a flow-through mode, endotoxin could be effectively removed from these proteins while maintaining very acceptable protein yields, and endotoxin levels were ultimately decreased to below 0.5 EU per microg of protein.

TL;DR: It appears that protein binds to MEP resin through both polar and hydrophobic interactions with some contribution of electrostatic interaction, which can be simultaneously reduced by arginine or urea.

TL;DR: Crystallization experiments demonstrated that the Fab fragments are of high quality to produce diffraction quality crystals suitable for X-ray crystallographic analysis.

TL;DR: Improvements to the previously reported ELP-intein purification system are discussed, which are able to reduce the required salt concentration by almost 4-fold, and the precipitation steps could be conducted at room temperature instead of 37 degrees C, resulting in a cheaper, gentler, and more scaleable purification method.

TL;DR: The chitinase producing Penicillium sp.

TL;DR: The use of monomeric Ocr (Mocr) as a complement to other fusion tags such as maltose-binding protein will provide greater flexibility in protein production and processing for a wide variety of protein applications.

TL;DR: The results suggest that the benefits of chaperone over-expression on the production of protein kinases in E. coli are indeed case-specific, and kinases behave in three different ways.

TL;DR: A dialysis reactor and a fully automated system for protein production using the dialysis cell-free synthesis method, validated to be able to simultaneously and automatically produce up to 96 proteins in yields of several milligrams with high well-to-well reliability, sufficient for structural and functional analyses of proteins.

TL;DR: Silver croaker chitinase was judged to be an enzyme that preferentially hydrolyzes the 2nd glycosidic link from the non-reducing end of (GlcNAc)n, which coincides well with the feeding habit of the silver croaker, which feeds mainly on these animals.

TL;DR: Kinetic analysis indicated that urea aided in suppressing insoluble precipitates, while arginine prevented formation of soluble oligomers produced by hydrophobic interaction with this combination system, the refolding yield of rhG-CSF could be increased 2-fold.

TL;DR: High-level extracellular production of Fusarium solani cutinase using a Pichia pastoris expression system was demonstrated for the first time and would be a promising alternative to many expression systems previously used for the large-scaleProduction of F. solanicutinase in Saccharomyces cerevisiae as well as Escherichia coli.

TL;DR: Two-dimensional fluorescence difference gel electrophoresis showed that differences in cell viability had more influence on the protein expression pattern than did the expression clone itself, indicating the validity of purification scheme.

TL;DR: The described approach is a promising method for producing large amounts of different recombinant BMPs that show potential for novel biomedical applications.

TL;DR: Stability in the presence of detergents, surfactants, metal ions and solvents, and sensitivity to various metal ions, additives and inhibitors make this keratinase suitable for industrial processes.

TL;DR: Heterologous expression in Aspergillus awamori of a betabeta-(1-4) endo-xylanase isolated from the whole-genome DNA sequence of A. clavatus is described along with a comprehensive biochemical and functional analysis of the enzyme, including substrate preference and hydrolysis patterns.

TL;DR: The synthesis of a human SR gene variant optimized for heterologous expression in Escherichia coli is reported and the expression and purification of active recombinant human SR are described, which may be of general interest to researchers wishing to express mammalian proteins in a bacterial system.

TL;DR: The authors cloned and expressed the two recombinant peptides as ketosteroid isomerase (KSI) fusion proteins inclusion bodies in Escherichia coli and subjected them to NMR structural studies to explore their structure-activity relationships.

TL;DR: A simple and efficient method for the expression of NS1 in Escherichia coli is described, which could potentially be used to develop monoclonal and bispecific antibodies for point of care diagnostics.

TL;DR: It is suggested that this expression system based on the trypanosomatid protozoan host Leishmania tarentolae has the potential for mass production of LMs for tissue engineering.

TL;DR: This protein can be used for immunoassays and it is suitable for vaccine candidate against A. hydrophila infection, which contributes pathogenicity of fish to humans.