Showing papers in "Protein Expression and Purification in 1996"

TL;DR: By constructing a basic gene unit encoding (GVGVP)10, it was possible to build concatemer genes with as many as 25 repeats of the monomer unit encoding a protein-based polymer with a molecular weight of greater than 100,000 Da to achieve purification of more complex polymers.

TL;DR: The methodology described will allow production of sufficient quantities of protein for basic structure/function studies including production of synthetic fibers and will have general utility in the controlled construction of repetitive proteins composed of identical or different repeat units.

TL;DR: The expression of the cytokine human Leukemia Inhibitory Factor (hu-LIF) in five of the most commonly used expression systems, namely in CHO, Sp2/0, MEL, COS, and insect cells, is summarized in conjunction with an outline of the principles and characteristics of each of these expression systems.

TL;DR: A new set of plasmids that can be used to clone and express foreign genes under the control of a baculovirus ie1 promoter are described and it is suggested that immediate-early bacULovirus vectors might be as useful as conventional baculinovirus expression vectors for producing biologically active eucaryotic secretory pathway proteins.

TL;DR: The two isoforms were characterized biochemically and all evidence, including that from analysis of the complex pre-steady-state kinetic behavior of the enzymes, indicates that the functional properties of the two isoenzymes are identical.

TL;DR: These findings are important because they demonstrate that culture conditions can significantly influence the concentration of a biologically active foreign protein secreted from plant cells into the media of suspension cultures.

TL;DR: Various genetic approaches to improve the stability of recombinant proteins will be discussed, including (i) choice of host cell strain, (ii) product localization, (iii) use of gene fusion partners, and (iv) product engineering.

TL;DR: The RAP fraction may define one or more functional forms of RNA polymerase II distinct from the activator-mediating holoenzyme, as well as two proteins with interesting connections to gene expression.

TL;DR: Pichia pastoris produced in high yields recombinant alpha-amylase that is similar with respect to structure and function to the enzyme purified from malt extracts, which greatly facilitates future mutational analysis of barley alpha- amylase in order to probe structure/function relationships.

TL;DR: The recombinant CDA has been purified to homogeneity by a rapid procedure consisting of heat inactivation followed by affinity chromatography and showed a specific activity of 105 U/mg, corresponding to about 88-fold purification with respect to the crude extract and was judged to be >98% pure by SDS-PAGE.

TL;DR: It is concluded that cysteines 135 and 175 are both necessary for efficient secretion of a biologically active N-terminal BPI fragment, presumably through the formation of a disulfide bond and cysteine 132 is responsible for dimer formation.

TL;DR: A method for the stepwise purification of bovine seminal plasma proteins aSFP, PDC-109, TIMP-2, RNAse, and BSP-30K/PDC-109 complex is presented.

TL;DR: A single-step method is devised that enables purification of HIV-1 recombinant reverse transcriptase directly from bacterial lysates in less than 2 h, and a crucial aspect of the procedure is the use of Tris buffer, a buffer that is normally incompatible in cation-exchange methods.

TL;DR: To generate sufficient quantity of TnT protein for antibody production, the corresponding cDNA was cloned and expressed in E. coli and two pairs of consecutive AGG codons were successively replaced with the major synonymous codon CGT by site-directed mutagenesis.

TL;DR: A recombinant form of hirudin (HIR), a potent thrombin inhibitor derived from the leech Hirudo medicinalis, was cloned and expressed in the methylotrophic yeast Pichia pastoris, offering an efficient system for production and purification of multigram quantities of biologically active rHIR for structure/function analyses.

TL;DR: A rapid, high-yield GRP94 purification procedure is reported that yields milligram quantities of homogeneous protein suitable for structural and biochemical analyses and indicates that the protein exists as a dimer of noncovalently associated subunits.

TL;DR: Refolding of the Link module, which occurred during the purification procedure, gave two species with different disulfide bond organizations that could be separated by high-performance liquid chromatography.

TL;DR: PDI-mediated rescue of proteins, perhaps assisted by chaperones and other foldases, may be important in vivo where insolubility is a common occurrence for newly synthesized polypeptides.

TL;DR: The first cloning, expression, and extensive purification of IpaB and IpaC fusion proteins from Escherichia coli for use in dissecting of the protein biochemistry of S. flexneri pathogenesis are described.

TL;DR: Evidence is presented that a 49-amino-acid region of the recombinant azurocidin protein is required for its secretion from insect cells, and it is shown that it retained the antimicrobial activity of the native neutrophil-derived molecule.

TL;DR: Methods reported here permit the efficient recovery of purified EBP which quantitatively binds EPO in solution as determined by high performance size exclusion chromatography and should facilitate the structural determination of the protein by NMR or crystallography and may serve as a guide for the refolding of other hematopoietic receptors.

TL;DR: It is concluded that BmNPV vectors can be used successfully for expressing chromosomal genes harboring multiple introns and retained the immunological and biological properties of the native peptide.

TL;DR: Characterization of affinity-purified antibodies revealed that there exists no cross-reactivity between the two fusion systems and that such antibodies indeed could be used for immunohistology, implications for the described system for large-scale functional analysis of cDNA libraries are discussed.

TL;DR: A synthetic gene encoding beta-cryptogein, a member of the elicitin family, has been cloned into a vector for expression by the methylotrophic yeast, Pichia pastoris and overexpressed a modified beta- cryptogein in a secreted form.

TL;DR: RC-RNase is proposed to have both ribonucleolytic and cytotoxic activity and a specific receptor on the tumor cell surface is suspected to be involved in the recognition and binding, and possibly entry of RC- RNase.

TL;DR: This represents the highest reported level of glycosylated, recombinant IFN expression in a stable mammalian system and is a significant advance in the large-scale production of these clinically important cytokines.

TL;DR: The present results suggest that a signal sequence is required for the efficient translocation of PA into E. coli periplasmic space.

TL;DR: Comparative ELISA data suggest that the 56.5-kDa protein served as a highly reactive antigen in detecting anti-HEV antibodies and may serve as a particularly useful antigen for both diagnostic and vaccine purposes.

TL;DR: Improved expression system gave higher yields of tryptophan synthase proteins and led to correspondingly high yields of purified alpha and beta2 subunits or alpha2beta2 complex from Salmonella typhimurium.

TL;DR: Evidence is presented that sulfotransferase alpha, but not beta, has 1 mol of PAP tightly bound per enzyme dimer, and the ability to utilize PAPS as a sulfate donor could be altered.