143results about "Metallothioneins" patented technology

Streptavidin-metallothionein chimeric proteins with biological recognition specificity in which the streptavidin moiety provides high affinity biotin binding and the metallothionein moiety provides a high affinity metal binding. The binding affinity of the streptavidin-metallothionein chimeric protein both for biotin and heavy metal ions allows specific incorporation into, conjugation with, or labelling of any biological material containing biotin with various heavy metal ions.

The present invention relates to a method of preparing heavy metal nanoparticles using a heavy metal-binding protein. More specifically, relates to a method for preparing heavy metal structures, comprising the steps of: culturing a microorganism transformed with a gene encoding a heavy metal-binding protein, in a heavy metal ion-containing medium, to produce heavy metal structures in the microorganism; and collecting the produced heavy metal structures, as well as nanoparticles of heavy metal structures prepared according to said method. Unlike prior methods of preparing quantum dots by physically binding metal materials, the quantum dots disclosed herein can be efficiently produced by expressing the heavy metal-binding protein in cells. In addition, the quantum dots are useful because they can solve an optical stability problem that is the shortcoming of organic fluorophores.

The present invention provides compositions and methods for regulating expression of heterologous nucleotide sequences in a plant. Compositions include a novel nucleotide sequence for a maize metallothionein 2 promoter for the gene encoding metallothionein. A method for expressing a heterologous nucleotide sequence in a plant using the promoter sequences disclosed herein is provided. The method comprises transforming a plant or plant cell with a nucleotide sequence operably linked to one of the promoters of the present invention.

The invention discloses a halostachys caspica metallothionein gene as well as a recombinant protein and application thereof. The halostachys caspica metallothionein gene has a base sequence shown by SEQ ID NO.1 in a sequence table and has a full length of 234bp, and is used for coding a protein consisting of 78 amino acids in an amino acid sequence shown by SEQ ID NO.2 in the sequence table. The invention also discloses a recombinant expression vector and an expression transformant containing the gene, and a preparation method and application of halostachys caspica metallothionein. According to the invention, a new metallothionein gene is obtained through in vivo cloning of a Xinjiang saline alkali plant namely halostachys caspica; the recombinant metallothionein obtained through production of a genetic engineering expression transformant contains 14 cysteines without a disulfide bond, and is rich in sulfydryl groups which can be chelated with heavy metals including zinc, copper, cadmium and the like and can clean free radicals and resist ionizing radiation or ultraviolet irradiation, so that the recombinant metallothionein has a wide application field.

The invention discloses a production method of metallothioneins of cordyceps sinensis, particularly comprising the following steps of: taking cordyceps sinensis; carrying out liquid culture (adding zinc sulfate for induction); filtering mycelia to obtain mycelia which are rich in the metallothioneins; adding a buffer solution, and grinding to disrupt cells; adding an organic denatured solution, and stirring to denature foreign proteins; centrifugally removing the foreign proteins; heating a supernatant for denaturing; centrifugally removing denatured proteins; fast cooling to room temperature; carrying out ultrafiltration concentration to obtain a concentrated solution; carrying out chromatography through G-50; monitoring an MT (Metallothionein) eluent through a sulfhydryl reagent method,and collecting the MT eluent; and carrying out vacuum freeze drying on the MT eluent to obtain MT freeze-dried powder. The invention has the advantages of easiness in operation, short production period, high yield of MT contained in the mycelia or sporocarps and factory production.

Metallothionein is extracted from pork liver by. taking porket as supply source, Intraperitoneal injecting with trace element zinc and inducing, Butchering after 5 times inducing, separating and extracting metallothionein which is well stability and high purity, with high output i.e. 2 to 3 times of extracted from rabbits. The pork liver metallothionein induced with zinc has advantages of low cost, good effect, high output, wide source of inducer, without side effect etc. And pig is a easy feeding animal, wide source, and the pork is edible after inducing to extract metallothionein, The products thereof adapted for anti-oxidation and anti irritability for avian and livestock's feedstuff.

This invention relates to a process for the manufacturing of a protein in mammalian cells cultured in a serum-free medium.

The invention relates to mitochondrial-targeting protein nanoparticles, and relates to an amino acid sequence, a coding nucleic acid sequence, a vector of the coding nucleic acid, host expression bacteria and other related information. Under the induction of metal ions, the self-assembled protein nanoparticles can be obtained by an escherichia coli expression system. The protein nanoparticles canbe applied in cancer diagnosis and treatment. Compared with conventional mitochondrial-targeting small molecules (TPP, MPP and the like), the protein nanoparticles can achieve properties of tumor enrichment, mitochondrial targeting, causing of increase of the ROS content in cells, induction of cell apoptosis, inhibition of tumor growth and the like.

The present invention relates to a mitochondria targeting peptide, a fusion protein in which the peptide is bound to the carboxyl terminus of a protein transduction domain, a fusion protein in which the peptide is bound to the carboxyl terminus of a protein transduction domain and an antioxidant is bound to the carboxyl terminus of the peptide, an antioxidant composition and a composition for preventing or treating Parkinson's disease including the fusion protein in which the antioxidant is bound, a recombinant polynucleotide in which a polynucleotide coding a protein transduction domain, a polynucleotide coding the peptide, and a polynucleotide coding an antioxidant protein are sequentially bound, to an expression vector including the polynucleotide, and to a transformed host cell including the expression vector.The mitochondria targeting peptide according to the present invention targets mitochondria with high efficiency not only when the peptide exists alone but also when the peptide is bound to a protein transduction domain and / or to an antioxidant. Further, the peptide has a small size and is thus a very suitable targeting carrier. The peptide becomes processed when introduced into mitochondria, and thus stably delivers the drug carried by the peptide.

Devices, such as solid supports having metal binding proteins, such as metallothionein proteins, bound thereto are disclosed for removing metals from substrates in need of having such metals removed therefrom. Specifically membranes having metallothionein proteins from the brine shrimp Artemia are disclosed for removing metals from liquid substrates. Associated methods for removing metals from substrates using metallothionein proteins are also disclosed.

Streptavidin-metallothionein chimeric proteins with biological recognition specificity in which the streptavidin moiety provides high affinity biotin binding and the metallothionein moiety provides a high affinity metal binding. The binding affinity of the streptavidin-metallothionein chimeric protein both for biotin and heavy metal ions allows specific incorporation into, conjugation with, or labelling of any biological material containing biotin with various heavy metal ions.

The invention relates to a method for inducing and extracting metallothionein from housefly larva, which comprises the following steps of: culturing 3-5-day housefly larva in a nonmetal culture dish by using a culture medium of a metal ion biological inductive agent of Cd2+, Zn2+ and Cu2+; and carrying out rinsing, sterilizing, low temperature and high speed homogenate, membrane filtration, chromatographic fractionation, elution and concentration, and vacuum freezing and drying on the housefly larva to obtain Zn / Cu metallothionein which has molecular weight of 5300-6500 dDa and contains 20-27 percent of cysteine and subtypes thereof, wherein the molecular structure of the Zn / Cu metallothionein takes less aromatic amino acids. The yield of the metallothionein in biologically induced larva protein can reach 1mg / g. The production method has low cost, high purity and high yield of metallothionein.

The invention discloses a ramie metallothionein gene and recombinant protein and an application thereof. The ramie metallothionein gene is a nucleotide sequence shown as the sequence table SEQ ID NO.1, and the ramie metallothionein is an amino acid sequence shown as the sequence table SEQ ID NO.2. A new metallothionein gene is obtained from ramie which is a herbaceous plant through in-vivo cloning, a recombinant expression engineering strain is constructed, and the recombinant metallothionein is obtained through expression and purification. The recombinant protein contains 12 cysteines (Cys),is abundant in mercapto groups that can chelate with cadmium and other heavy metal ions, and has wide application prospects in the fields of preparation of a heavy metal chelating antidote, chemical or medicine or heavy metal polluted cultivated land and pollution treatment of agricultural non-point sources.

Devices, such as solid supports having metal binding proteins, such as metallothionein proteins, bound thereto are disclosed for removing metals from substrates in need of having such metals removed therefrom. Specifically membranes having metallothionein proteins from the brine shrimp Artemia are disclosed for removing metals from liquid substrates. Associated methods for removing metals from substrates using metallothionein proteins are also disclosed.

The invention discloses Yunnan red pear-1 metallothionein gene PyMT1 and a prokaryotic expression vector thereof, and the prokaryotic expression vector is a prokaryotic expression vector which contains the Yunnan red pear-1 metallothionein gene PyMT1. According to the invention, PyMT1 gene is cloned from Yunnan red pear-1; the over expression of the gene in bacterial cells is controlled by T7 promoter and lactose operon with the induction of IPTG; PyMT1 protein is synthesized and secreted into cytoplasm; the expression level is detected by SDS-PAGE; test results show that the expression level of the PyMT1 protein reaches a maximum when the expression is inducted by 1mM IPTG at 37 DEG C for 4 hours and when the expression is inducted by 0.5 mM IPTG at 37 DEG C for 4 hours; the vector adopted by the invention can successfully express a lot of Yunnan red pear-1 metallothionein; the test results also demonstrates that the Yunnan red pear-1 metallothionein gene PyMT1 of Yunnan red pear has a typical function of heavy metal ion tolerance of the metallothionein gene, which lays a foundation for further research of PyMT1 functions and the applications of the metallothionein gene in microbial tolerance to heavy metal.

The invention discloses a method which is feasible in pharmaceutical industry for efficient secretion expressing, preparing and purifying of gene recombinant human metallothionein or analogues thereof. The method is characterized by introducing exogenous genes containing dual-promoters into relevant host yeast cells in transformation or double crossover recombination manner by using plasmid vector pPICZ alpha-MT of exogenous metal sulfur protein genes which contain methanol response element AOX and contain or not contain metal response element MRE to construction expression engineering cells containing the dual-promoters in the methanol yeast host cells. According to the invention, several problems with traditional preparation method of metallothionein, such as high production cost, slaughter and organ harvesting to large number of animals, virus pollution derived from animal, endotoxin pollution derived from bacterial, immunogen of non-humanized protein and the like, are avoided effectively,.

The invention belongs to the bio-pharmaceuticals field, in particular to a method for preparing metallothionein by taking the liver of sturgeon as raw material. The method mainly comprises steps: sturgeons fed with feed added with inducer-zinc acetate by humans are slaughtered to take livers thereof; liver homogenate buffer solution is used for homogenizing and centrifuging, glucan and iminodiacetic acid mixture chromatography column chromatography collects chromatography liquid, and then the metallothionein is obtained by vacuum cooling drying. The liver of sturgeon is rich in metallothionein and is 100 times of liver content of other animals. The method is used for extracting the metallothionein, which can reduce the cost by 30%, increases yield by 40% and quality, and the purity the metallothionein reaches more than 99%. The metallothionein is expensive, has obvious physiological function and extensive use. Extracting metallothionein by using the liver of sturgeon has wide market prospect.

The invention provides a gene cloning method, expressing method and application of psychrophilic yeast metallothionein. Metallothionein genes are extracted, a primer F1 and a primer R1 are designed, subjected to PCR amplification and then connected with an expression carrier, and a recombinant expression carrier is constructed; the recombinant expression carrier is converted into an expression strain, and a recombination expression transformant is constructed; the expressing and purifying method of recombinant protein is built. The nucleotide sequence and the coded amino acid sequence of the metallothionein genes are shown in the sequence table SEQ ID NO:1 and the sequence table SEQ ID NO:2. The novel metallothionein obtained through cloning comprises 16 pieces of cysteine, multiple metal and heavy metal ions can be chelated through rich sulfydryl, and the novel metallothionein has broad prospects in the medicine field, the food health caring field, the cosmetic additive field, the genetic engineering reagent field, the chemical engineering field, the environmental protection field, the animal field, the agriculture field and the like.

The present invention provides compositions and methods for regulating expression of heterologous nucleotide sequences in a plant. Compositions include a novel nucleotide sequence for a promoter for the gene encoding metallothionein. A method for expressing a heterologous nucleotide sequence in a plant using the promoter sequences disclosed herein is provided. The method comprises transforming a plant or plant cell with a nucleotide sequence operably linked to one of the promoters of the present invention.

The invention relates to an industrial method for extracting and purifying zinc metallothionin from rabbit liver. It comprises following steps: inducing, coarse extracting, chromatography purifying; inducing adult rabbit for 5- 7 times through belly hypodermic injection, the zinc injection dosage is increased by 10- 250 mg for every kg rabbit gradually; stirring rabbit liver with homogenate liquor, adding denatured liqour and stirring, freezing and centrifuging, collecting supernatant -1, heating fastly, colling fastly to normal temperature, freezing and centrifuging again, getting supernatant -2, condensing through ultra- filtering membrane, getting supernatant -3 for chromatography in gel medium, desalting collecting liquid and freeze drying. The invention is characterized by higher productivity of Zn- MT, low extracting cost, high purity, simple preparation process and convenient extracting method.

The present invention concerns a method for modifying the growth characteristics of plants by modulating expression in a plant of a nucleic acid sequence encoding a metallothionein and / or modulating activity in a plant of a metallothionein. The invention also relates to transgenic plants having modified growth characteristics, which plants have modulated expression of a nucleic acid encoding a metallothionein.

The invention relates to an African swine fever virus P22 protein nanoparticle and a preparation method and application thereof, the protein nanoparticle is formed by self-assembly of a protein monomer, and the protein monomer is a fusion protein sequentially containing metallothionein, glutathione thiotransferase and P22 protein from amino acid to a carboxyl terminal. The GSTP1-MT3 protein and the P22 antigen of the African swine fever virus are subjected to fusion expression, and in a pichia pastoris expression system, protein nanoparticles can be spontaneously formed through induction of ferrous ions. Immunogenicity determination is carried out in a mouse body, and the P22 protein nanoparticle can cause very strong immune response and has great potential to be developed into a new safeand effective African swine fever virus vaccine.

A device for the removal of heavy metals and heavy metal complexes, such as thimerosal, from medications is provided wherein the device comprises a dosing device or a solid support having at least one substantially purified metallothionein protein associated therewith. Additionally, methods to remove thimerosal from medications are provided.

A device for the removal of heavy metals and heavy metal complexes, such as thimerosal, from medications is provided wherein the device comprises a dosing device or a solid support having at least one substantially purified metallothionein protein associated therewith. Additionally, methods to remove thimerosal from medications are provided.

The present invention provides compositions and methods for regulating expression of heterologous nucleotide sequences in a plant. Compositions include a novel nucleotide sequence for a maize metallothionein promoter for the gene encoding metallothionein. A method for expressing a heterologous nucleotide sequence in a plant using the promoter sequences disclosed herein is provided. The method comprises transforming a plant or plant cell with a nucleotide sequence operably linked to one of the promoters of the present invention.

The invention discloses application of metallothionein-1 in the preparation of drugs for inhibiting rheumatoid arthritis. The metallothionein-1 inhibits the differentiation of Th17 cells, promotes the differentiation of regulatory T cells, regulates the balance of Th17 / Treg under the state of CIA, and participates in the generation and development of rheumatoid arthritis; and the metallothionein-1 inhibits the expression of various inflammatory factors related to rheumatoid arthritis by a negative feedback regulation mechanism, and promotes the expression of immune suppression factors (IL-10 and TGF-beta) by a positive feedback regulation mechanism, and therefore, inflammatory response and pathologic change of rheumatoid arthritis are reduced, the process of the rheumatoid arthritis is delayed, and clinical evidence and a drug target are provided for the diagnosis and treatment of autoimmune diseases related to rheumatoid arthritis.

In order to provide a composition for treating, improving, or preventing pain, it has been found out that a metallothionein protein is expressed in a normal peripheral nerve, and that the expression is not observed in a peripheral nerve producing pain. Moreover, it has been found out that administering metallothionein to rats having neuropathic pain suppresses the pain in the rats.

The present invention provides compositions and methods for regulating expression of heterologous nucleotide sequences in a plant. Compositions include a novel nucleotide sequence for a promoter for the gene encoding metallothionein. A method for expressing a heterologous nucleotide sequence in a plant using the promoter sequences disclosed herein is provided. The method comprises transforming a plant or plant cell with a nucleotide sequence operably linked to one of the promoters of the present invention.