175 results about "Thioredoxin" patented technology

An expression system which provides heterologous proteins expressed by a non-native host organism but which have native-protein-like biological activity and/or structure. Disclosed are vectors, expression hosts and methods for expressing the heterologous proteins. The expression system involves co-expression of protein factor(s) which is/are capable of catalyzing disulphide bond formation and desired heterologous protein(s). The expression system is presented using yeast cells as the preferred host, protein disulphide isomerase (PDI) and thioredoxin (TRX) as the preferred examples of the protein factors and HCV-E2715 envelope glycoprotein and human FIGF as the preferred examples of the heterologous proteins.

The present invention is directed to prodrugs of hydroxamic acid based histone deacetylase (HDAC) inhibitors, e.g., suberoylanilide hydroxamic acid (SAHA). The prodrugs are acylated derivatives having increased aqueous solubility and cellular permeability as compared with the free hydroxamic acid, and are useful for inhibiting HDACs, and for selectively inducing terminal differentiation, cell growth arrest and/or apoptosis of neoplastic cells, thereby inhibiting proliferation of such cells. Thus, the prodrugs of the present invention are useful in treating a patient having a tumor characterized by proliferation of neoplastic cells. The prodrugs of the invention are also useful in the prevention and treatment of thioredoxin (TRX)-mediated diseases, such as autoimmune, allergic and inflammatory diseases, and in the prevention and/or treatment of diseases of the central nervous system (CNS), such as neurodegenerative diseases.

An intracellular selection system allows screening for peptide bioactivity and stability. Randomized recombinant peptides are screened for bioactivity in a tightly regulated expression system, preferably derived from the wild-type lac operon. Bioactive peptides thus identified are inherently protease- and peptidase-resistant. Also provided are bioactive peptides stabilized by a stabilizing group at the N-terminus, the C-terminus, or both. The stabilizing group can be a small stable protein, such as the Rop protein, glutathione sulfotransferase, thioredoxin, maltose binding protein, or glutathione reductase, an α-helical moiety, or one or more proline residues.

The invention relates to methods for monitoring patient response to histone deacetylase inhibitors (e.g., suberoylanilide hydroxamic acid (SAHA)) or other therapeutic agents by measuring the level of thioredoxin in body fluids, tissues, and/or cells, such as peripheral blood mononuclear cells, plasma, or serum. The invention also relates to methods of monitoring and/or assisting with the diagnosis of a wide variety of thioredoxin-related diseases and conditions, such as inflammatory diseases, allergic diseases, autoimmune diseases, diseases associated with oxidative stress or diseases characterized by cellular hyperproliferation.

Novel DNA constructs and host cells comprising the same are disclosed. DNA constructs comprise a transcription unit (e.g. operon) comprising DNA sequences encoding for ribonucleotide reductase and thioredoxin. In preferred embodiments, constructs further comprise DNA sequences encoding for thymidylate synthase and/or transcription units comprising sequences encoding for uridine kinase preferably together with dCTP deaminase. In particularly preferred embodiments, host cells comprising constructs having all of the above characteristics wherein the host cell displays repressed or no uracil DNA glycosylase activity. This may be achieved by removal of the host cell ung gene. Use of host cells in the manufacture of pyrimidine deoxyribonucleotides e.g. thymidine is also disclosed.

An intracellular selection system allows concurrent screening for peptide bioactivity and stability. Randomized recombinant peptides are screened for bioactivity in a tightly regulated expression system, preferably derived from the wild-type lac operon. Bioactive peptides thus identified are inherently protease- and peptidase-resistant. Also provided are bioactive peptides stabilized by a stabilizing group at either the N-terminus, the C-terminus, or both. The stabilizing group can take the form of a small stable protein, such as the Rop protein, glutathione sulfotransferase, thioredoxin, maltose binding protein, or glutathione reductase, or one or more proline residues.

The present invention generally relates to methods of preselecting patients for treatment with an anti-VEGF therapy, anti-HIF-1 therapy or anti-thioredoxin therapy. Aspects of the invention combine methods of dynamic contrast enhanced-MRI and diffusion weighted-MRI for the detection of tumor histology. The methodology disclosed herein detects tissue blood volume, tumor vascularity, and abnormal capillary permeability, thereby determining tumor vascularity to determine whether a patient should be administered such therapy.

The invention relates to application of protein and proteome in preparation of a liver cirrhosis diagnostic reagent. The protein comprises Lumican, Tetranectin, Pro-platelet basic protein (PBP), Pigment epithelium-derived factor (PEDF), Insulin-like growth factor binding protein 3 (IGFbp-3), Sex hormone-binding globulin (SHBG), Thioredoxin and composition of parts in the liver cirrhosis diagnostic reagent. Multiple proteins related with health and disease states are identified and evaluated in large range by adopting a proteome technology, the screened biomarkers for identifying hepatitis and liver fibrosis are used for predicting the degree of hepatitis and liver fibrosis and can be used for preparing a kit for diagnosing liver cirrhosis and liver cirrhosis level, and the protein markers identify the sensitivity of liver cirrhosis and degree thereof; and by combining the optimal conditions of various protein markers, the sensitivity can reach 89 percent, the specificity reaches over 90 percent, and the sensitivity and the specificity are higher than those of any known detection method.

The invention discloses a preparation method of a fusion protein Trx-TAT-hMsrA and application thereof to an aspect of nerve cell protection. The fusion protein Trx-TAT-hMsrA comprises a TAT protein transduction structure domain, thioredoxin Trx and anthropogenic methionine sulfoxide reductase hMsrA. The fusion protein of the invention has various biological effects for reducing the oxidization of functional protein methionine residues, treating oxidative stress injury, relieving the relevant nerve cell pathological change of parkinsonism, preventing and treating calcium ion overloading and the like, simultaneously has good capability for entering nerve cells through the epicyte, and belongs to novel biotechnological medicine candidates with good druggability.

The invention belongs to the technical field of biomedicine and provides a cyanea capillata thioredoxin. Currently, researches and reports related to jellyfish thioredoxin are not seen yet. The cyanea capillata thioredoxin provided by the invention has protein formed by an amino acid sequence shown in SEQ ID NO: 2. The invention further provides a coding gene of the cyanea capillata thioredoxin, wherein the coding gene has a nucleotide sequence shown in SEQ ID NO: 1. Meanwhile, the invention further provides application of the cyanea capillata thioredoxin and the coding gene of the cyanea capillata thioredoxin in preparation of anti-oxidation drugs, anti-radiation drugs, skin protectants, food preservatives, anti-aging drugs and the like. The cyanea capillata thioredoxin and the coding gene of the cyanea capillata thioredoxin have greater clinical application values.

The invention discloses a fusion protein for Micrococcus pyogenes surface protein, and also discloses the method for preparing the same as well as its usage. The fusion protein is fused by Micrococcus pyogenes fnb and thioredoxin Trx on expression carrier, and the process comprises preparation of Micrococcus pyogenes surface protein fnb coding gene, colony of fnb coding gene into expression carrier and protein fusion. The fusion protein is soluble protein and retains immunity of natural protein. It is detected that said fusion protein is characterized by good response for humoral immunity and cell immunity, enlargement of protection range of protein vaccine and prevention of infection caused by most of Micrococcus pyogenes.

Disclosed herein are nucleic acid constructs and expression vectors for enhancing the production of recombinant polypeptides/proteins, and methods for the massive production of recombinant polypeptides/proteins, in which a first nucleic acid sequence encoding thioredoxin and a second nucleic acid sequence encoding hemoglobin are cloned into a host cell, thereby enhancing the capability of the thus formed recombinant host cell in producing a selected gene product, and thereby assisting said recombinant host cell in relieving intracellular stress due to the overproduction of said gene product.

The invention discloses a method for preparing recombined human insulin growth factor-1(IGF-1) fused protein, which belongs to the field of biological medicine field in biological technique. The invention adopts the genetic engineering fungus fermentation method for production: a. designing and synthesizing recombining human IGF-1 fused protein gene; b. constructing an expression vector of the recombined human IGF-1 fused protein; c. utilizing the expression vector to convert the host to construct the genetic engineering fungus; and d. utilizing the genetic engineering fungus to ferment the recombined human IGF-1 fused protein. In the method, according to the characteristics that the first amino acid of the natural IGF-1 is glycine(GLy, G), a leading peptide is added before the natural IGF-1, and a hydroxylamine specific cracking part(Asn-Gly peptide bond) is designed between the leading peptide and the natural IGF-1, thereby making the expression products stable and lowering purification cost. In the method, the fused protein in serial expression is used, wherein the N end is thioredoxin, and the C end is IGF-1, so that the expression products are more stable, and the separation method is simple and cheap. The method has wide application prospect.

The current invention provides methods for producing a polypeptide as inclusion bodies in bacterial host cells. The present methods are carried out by forming a gene construct comprising the genetic sequence encoding a polypeptide operatively linked to that of an inclusion partner protein, such as E. coli thioredoxin or a modified E. coli thioredoxin, such that host cells comprising the gene construct produce the polypeptide as intracellular inclusion bodies. The methods of the present invention facilitate the rapid isolation and purification of recombinant proteins. In addition, the present methods may be useful for producing polypeptides or proteins which are small and are typically difficult to express, as well as those proteins that are toxic to host cells such as E. coli. The present invention also provides plasmids, vectors and host cells to be used in the present invention for production of polypeptides, and methods of production of polypeptides using these vectors and host cells. The invention further provides methods for producing protein molecular weight ladders for use in protein gel electrophoresis, as well as proteins and protein molecular weight ladders produced by these methods.

The invention relates to the technical field of biomedical engineering, and relates to a method for preparing human parathyroid hormone (1-34) through fermenting and purifying human parathyroid hormone 1-34 recombinant engineering bacteria constructed by a gene engineering method. The method provided by the invention comprises the following steps: fusing and expressing the genes for coding human parathyroid hormone and a thioredoxin label, and further constructing and finishing genetic engineering bacteria capable of highly expressing human parathyroid hormone; adding IPTG (isopropyl beta D thiogalactopy ranoside) and triton X-100 in the fermentation process by adopting a chemosmosis fermentation technology so as to improve the release of proteins extracellularly in the fermentation process, which is good for the continuous synthesis of proteins, and avoiding the attack of endoproteinase, thus protein high-yield extracellular accumulation is promoted; and simultaneously carrying out heat treatment on a fermentation liquid when the fermentation reaches the final point aiming at the heat stability of thioredoxin fusion, thus initially purified target proteins are directly recycled and obtained from a fermentation liquor supernatant after centrifugation.

The invention relates to a method for constructing porcine fmd recombinant immune complex peptide and an application thereof, which belongs to the technical field of biological engineering. The immune multi-peptide adjuvant is fused with aphthovirus novel auxiliary T cell epiposition (I-S-I-S-E-I-K-G-V-I-V-H-K-I-E-T-I-L-F) and Bursin mimic peptide (T-P-N-L-K-H-G) by a flexible Linker. A fusion gene is inserted into an expression carrier to be converted into colon bacillus, gene engineering strains which express the porcine fmd recombinant immune complex peptide with high efficiency are obtained, and the fusion protein of the porcine fmd recombinant immune complex peptide is obtained after liquid culture and purification; the thioredoxin in the fusion protein is removed by enterokinase with His label on the N end, and the single porcine fmd recombinant immune complex peptide can be obtained after affinity chromatography purification with very high sample purity. The recombinant immune complex peptide can serve as the novel immune multi-peptide adjuvant of the foot-and-mouth disease vaccine. Animal immune experiments prove that the method has high safety but no toxicity and side effect, and can effectively enhance the immunity of organisms and cells and the immunity level of body fluid.

This invention provides a method for preparing recombinant human parathyroid hormone 1-34 (rhPTH (1-34)). The method comprised: (1) expressing with an expression vector capable of expressing a fusion protein with an amino acid sequence (from N-terminal to C-terminal) containing thioredoxin, (His) 6, enterokinase recognition site and parathyroid hormone 1-34 peptide; (2) purifying the fusion protein by nickel ion complexation affinity chromatography; (3) cutting the purified fusion protein with enterokinase to release the rhPTH (1-34) from the fusion protein. By this method, rhPTH (1-34) can be purified rapidly, simply and efficiently, and the recovery yield is largely increased.

The invention relates to the fields of gene recombination and prokaryotic expression of proteins, and concretely relates to a double-solubilized expression tag sequence formed through fusing Escherichia coli thioredoxin TrxA and a small ubiquitin-like modifier SUMO, and an application thereof in a prokaryotic expression vector. An SUMO and TrxA double-solubilized expression tag has a base sequencerepresented by SEQ ID NO:1 in a sequence table. The sequence is applied to the upstream of the foreign protein encoding gene of the prokaryotic expression vector, and participates in fusion expression as a solubilizing tag together with a foreign gene in order to significantly promote the solubility of the expressed foreign protein and improve the recovery efficiency of a soluble foreign protein.

This invention provides a nucleic acid which encodes a soluble polypeptide which comprises the extracellular domain of a gonadotropin receptor and thioredoxin, wherein the soluble polypeptide is capable of binding to the gonadotropin. This invention also provides the polypeptides encoded by these nucleic acids and methods to produce the polypeptides. This invention also provides methods of preventing or terminating pregnancy, preventing or treating cancer, and decreasing androgen and estrogen production.

The present invention provides a kind of fusion protein, which possesses thioredoxin sequence and parathormone peptide 1-34 located in the downstream of the thioredoxin. Preferably, the fusion protein possesses also connecting peptide containing proteolytic enzyme recognizing site and located between the thioredoxin and the parathormone peptide 1-34. From the fusion protein, human parathormone peptide 1-34 may be prepared.