76 results about "Chaperone (protein)" patented technology

The present invention relates to an RNA encoding a therapeutic protein, in particular a collagenase, growth factor, cytokine, receptor, chaperone or signal transduction inhibitor. In particular, the present invention relates to RNA suitable for treatment of wounds, specifically for promoting wound healing. The present invention concerns such RNA as well as pharmaceutical compositions and kits and combinations comprising the RNA. Furthermore, the present invention relates to the RNA, pharmaceutical compositions, kits as disclosed herein for use in the treatment of wounds, specifically for promoting wound healing.

The invention relates to a prokaryotic expression vector and application thereof. The vector comprises: (a), a molecular chaperone for promoting the soluble expression of target protein or/and the correct folding of disulfide bonds; and (b), an encoding nucleotide sequence of intein which can be broken on peptide bonds at a C terminal. In the vector, the high-efficiency soluble expression of the target protein is promoted by the molecular chaperone, so that the target protein maintain natural bioactivity, and the self-shearing action of the intein is induced effectively under a certain condition to separate the target protein from fusion protein, and the target protein is separated and purified again to obtain the target protein with natural bioactivity economically and efficiently. The invention also relates to host cells containing the vector and application of the vector in the recombinant expression of bioactivity protein.

The invention discloses novel phosphorylation sites identified in signal transduction proteins and pathways, and provides phosphorylation-site specific antibodies and heavy-isotope labeled peptides (AQUA peptides) for the selective detection and quantification of these phosphorylated sites/proteins, as well as methods of using the reagents for such purpose. Among the phosphorylation sites identified are sites occurring in the following protein types: adaptor/scaffold proteins, adhesion/extracellular matrix protein, apoptosis proteins, calcium binding proteins, cell cycle regulation proteins, chaperone proteins, chromatin, DNA binding/repair/replication proteins, cytoskeletal proteins, endoplasmic reticulum or golgi proteins, enzyme proteins, G/regulator proteins, inhibitor proteins, motor/contractile proteins, phosphatase, protease, Ser/Thr protein kinases, Protein kinase (Tyr)s, receptor/channel/cell surface proteins, RNA binding proteins, transcriptional regulators, tumor suppressor proteins, ubiquitan conjugating system proteins and proteins of unknown function.

The invention relates to the technical field of biological engineering, in particular to a process method for effectively expressing a recombinant human coagulation factor VIII by utilizing mammalian cell culture. The method specifically comprises the following steps: adding an exogenous angiohemophilia factor to cell culture liquid for expressing the recombinant human coagulation factor VIII, and controlling an active ratio of the angiohemophilia factor to the human coagulation factor VIII in the cell culture liquid to be 1-10: 1 in the culture process. According to the invention, by utilizing a VWF (Von Willebrand Factor) molecule partner, which is necessary in an expression process of the recombinant human coagulation factor VIII, the newly generated human coagulation factor VIII is stabilized, the accumulating effect of the human coagulation factor VIII is improved, the technical difficulty is reduced, and the expression effect of a target protein is improved.

The present invention relates to a chimeric protein containing an intramolecular chaperone (IMC) like sequence linked to a target protein, preferably an insulin precursor. The present invention also relates to a process for obtaining a correctly folded insulin-precursor-containing chimeric protein, comprising, inter alia, contacting an incorrectly folded chimeric protein containing an IMC like sequence linked to an insulin precursor with at least one chaotropic auxiliary agent. The present invention further relates to an assay for screening an amino acid sequence for the ability to improve folding of an insulin precursor using a chimeric protein containing an IMC like sequence linked to an insulin precursor.

The present invention provides methods and compositions for enhancing channel activity to the mutant cystic fibrosis trans-membrane conductance regulator protein (CFTR). The compositions of the invention comprise polypeptides containing CFTR sub-domains that are designed to mimic the folding defect of the full length mutant CFTR proteins, resulting in competitive binding to cytoplasmic chaperones such as Hsc/Hsp7O and Hdj2. The methods of the invention comprise transduction, or recombinant expression, of CFTR polypeptides in a cell expressing mutant CFTR. The presence of the CFTR polypeptide results in a dominant effect whereby the CFTR polypeptide competes with the endogenously expressed mutant CFTR for binding to cytoplasmic chaperones such as Hsc/Hsp70 and Hdj2. Mutant CFTR proteins include, but are not limited to, DeltaF508 CFTR. The present invention is based on the discovery that reduced binding of cytoplasmic chaperones to the endogenous DeltaF508 CFTR, mediated by the presence of CFTR polypeptides, results in restoration of plasma membrane localization and channel activity. The methods and compositions of the invention can be used to restore channel activity in cystic fibrosis subjects carrying genetic defects in the CFTR gene, such as for example, DeltaF508 CFTR.

The invention discloses a preparation method of abyss sea cucumber sourced superoxide dismutase Cu, ZnSOD and application thereof. The method includes: extracting total RNA of Paelopatides sp., reversely transcribing into cDNA through RT-PCR, acquiring a gene sequence PaSOD coding superoxide dismutase through PCR amplification, establishing prokaryotic expression recombinant plasmid in pCold II vector, and guiding into chaperone Competent Cell pG-KJE8/BL21 for recombinant protein soluble expression. Recombinant protein prepared through the method overcomes the defects that raw material sourcesare difficult to obtain and protein renaturation recycling process is complex and troublesome, purified protein is high in purity and vitality, wide in temperature suiting range and capable of resisting digestion by high-concentration digestive enzyme, and a foundation is laid for widely applying the protein in the field of biology, food, medicine and cosmetology.

The invention discloses a recombinant expression vector for improving soluble expression of pathogenesis-related proteins in Mongolia astragaloside. The recombinant expression vector is a recombinantpET30a-skp-AmPR-10 constructed by using pET30a as a vector and modifying AmPR-10 by utilizing an escherichia coli molecular chaperone skp. According to the invention, by a method of replacing the vector and carrying out molecular chaperone modification, the soluble expression of target proteins is improved so as to fulfill the aims of saving cost and improving yield; the exogenously expressed AmPR-10 is further subjected to activity measurement so as to show that the exogenously expressed proteins also have the nuclease activity and provide reference strategies for in-vitro expression of otherhomologous related proteins; and the activity may have the important significance for researching the RNA virus resistance ability of radix astragali.

The invention discloses a method for synergistically and efficiently preparing a protein-based nano-emulsion by taking polyhydroxy alcohol as a molecular chaperone and the prepared protein-based nano-emulsion. The method comprises the following steps of dissolving protein in a polyhydroxy alcohol solution, performing uniform mixing to obtain a protein dispersion solution, performing mixing and homogenizing with an oil phase to obtain a crude emulsion, and performing emulsifying to obtain the protein-based nano-emulsion. According to the method, the functional characteristic that a polyhydroxycompound can be used as an active protective agent of enzyme or other functional proteins is utilized, and the protein and polyhydroxy alcohol are premixed, so that the original conformation of the protein in the subsequent high-energy emulsification process is effectively protected; and mixing with an oil phase is performed, and high-pressure homogenization is performed to obtain nano-scale oil-in-water emulsion with uniform particle size and good stability. The protein-based nano-emulsion provided by the invention is simple in materials, is food-grade, can be directly obtained through a convenient homogenizing means, and has an excellent protection or controlled release effect on the oil phase, so that the protein-based nano-emulsion has a wide application prospect in the fields of dailychemical products, foods, medicines and the like.

The invention discloses 432 novel acetylation sites identified in signal transduction proteins and pathways underlying human protein acetylation signaling pathways, and provides acetylation-site specific antibodies and heavy-isotope labeled peptides (AQUA peptides) for the selective detection and quantification of these acetylated sites/proteins, as well as methods of using the reagents for such purpose. Among the acetylation sites identified are sites occurring in the following protein types: Acetyltransferases, Adaptor/Scaffold proteins, Actin binding proteins, Adhesion proteins, Apoptosis proteins, Calcium-binding proteins, Cell Cycle Regulation proteins, Cell Surface proteins, DNA binding proteins, DNA replication proteins, Channel proteins, Chaperone proteins, Cellular Metabolism enzymes, Cytoskeletal proteins, DNA repair proteins, Endoplasmic reticulum proteins, Enzyme proteins, G protein and GTPase Activating proteins, Guanine Nucleotide Exchange Factors, Helicase proteins, Isomerase proteins, Extracelluar matrix proteins, Hydrolases, Ligase proteins, Lipid kinases, Inhibtor proteins, Lipid Binding proteins and Lyases.

The invention belongs to preparation of a new gene, construction of engineering bacteria, and particularly relates to a pyranose oxidase gene constructed from molecular chaperone, protein, phchia pastoris and preparation and application of the pyranose oxidase gene. Genes Ero I, PDI, CNE1, SEC53 and the like in pichia pastoris are respectively cloned and connected to series derivative plasmids ofpichia pastoris expression vectors pPICZ and the like for connection, and the genes and P20 are subjected to coexpression to realize conversion into Pichia pastoris GS115 bacterial strains to be coated to YPD flats being different in bleomycin concentration gradients for inspection of effects. Through overexpression of molecular chaperone and optimization of the fermentation condition, and throughscreening and identifying, a bacterial strain with enhanced secretory expression pyranose oxidase than an original bacterial strain is obtained. For the pyranose oxidase expressed by the bacterial strain, the enzyme activity in a 10L fermenter can achieve 405U/mL, and compared with the bacterial strain without overexpression of the molecular chaperone, the enzyme activity is improved by twice, sothat a favorable foundation is established for large-scale production of the pyranose oxidase.

The disclosure relates to a multi-epitope fusion protein as well as to its use as calibrator and/or control in an in vitro diagnostics immunoassay for detecting HCV core antigen. The multi-epitope fusion protein has two to six different non-overlapping linear peptides present in the amino acid sequence of hepatitis C virus (HCV) core protein, wherein each of the peptides is separated from the other peptides by a spacer consisting of a non-HCV amino acid sequence and having a chaperone amino acid sequence. No further HCV specific amino acid sequences are present in the polypeptide. A further aspect relates to a reagent kit for detecting HCV core antigen containing said multi-epitope fusion protein as calibrator or control or both.

The invention discloses application of a PGB protein in construction of a fusion protein expression vector with a chaperone-like protein effect. The fusion protein expression vector with the chaperone-like protein effect is successfully constructed by utilizing the PGB protein. For the application, by constructing a v chaperone-like protein contained pET-PGB3 expression vector and testing a targetprotein fusion protein expression vector and a target protein contained pET control vector, through contrast tests of target protein induction expressions in different recombinants, discovered that PGB has a novel chaperone-like protein function for improving the soluble expression efficiency of the target protein, so that a useful tool is provided to scientific research and industrial productionof protein expression.

The invention provides achaperone/usher family polymer comprising at least one chaperone/usher family polypeptide monomer, wherein said at least one chaperone/usher family polypeptide monomer comprises an exogenous bioactive sequence.

The invention relates to the technical field of molecular biology, in particular to a method for efficiently expressing structural proteins of Senecavirus A. According to the method, firstly, prokaryotic expression codon optimization is carried out on genes for coding three structural proteins VP0, VP3 and VP1 of the Senecavirus A, and a single plasmid for simultaneously carrying out soluble expression of the three structural proteins in escherichia coli is obtained by means of a small ubiquitin-like fusion protein; secondly, molecular chaperone plasmids and plasmids of the structural proteins of the Senecavirus A are transferred into the escherichia coli, and thus, the soluble expression of the structural proteins of the Senecavirus A is further improved, and the problem of non-uniform expression of target proteins is solved, and the obtained target protein accounts for about 25% or more of the total protein of bacteria. A Senecavirus subunit vaccine prepared from the three structural proteins obtained by expression can stimulate a pig body to generate a high-level neutralizing antibody, and has a good protection effect on a Senecavirus epidemic strain.

A portable rapid test strip for syphilis for household use is disclosed. Syphilis antigens TP17 and TP47 are adopted to construct an escherichia coli expression vector. Through codon optimization, carrying of purification tags, construction of the syphilis antigen expression vector and addition of a molecular chaperone signal peptide having a molecular weight of 12 KD, a target gene can be expressed efficiently in vitro and the exogenous target gene in a form of a soluble protein is secreted into periplasm, and therefore subsequent purification of target antigens is convenient. Through codon preference of the escherichia coli, the two antigen genes of the syphilis are optimized and artificially synthesized, efficient expression of the syphilis antigens in the escherichia coli can be achieved, and soluble expression can be achieved through controlling expression conditions, thus avoiding tedious and inefficient denaturation and renaturation processes of inclusion bodies. In terms of clone strategies, the C ends of the antigens carry the purification tags, and therefore product purification steps are simplified. The test strip is rapid, simple, economical, easily popularized, easily available in materials, and not restricted by experiment conditions.

The invention discloses a recombinant vector and an expression and purification method of heat-labile UNG fusion protein. The method comprises the following steps: cloning a gene sequence of Cod UNG to a pCold-sumo plasmid to construct a recombinant plasmid pCold-sumo-Cod UNG, transforming the recombinant plasmid into an Escherichia coli BL21 (DE3) competent cells, simultaneously transforming a plasmid pGTf2 which expresses chaperone into the competent cells, and carrying out low-temperature-induced expression to obtain the soluble sumo-Cod UNG fusion protein. According to the invention, the problem of insoluble Cod UNG expression products is successfully overcome.