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46 results about "Thionin" patented technology

Thionins (without an e) are a family of small proteins found solely in higher plants. Typically, a thionin consists of 45–48 amino acid residues. 6–8 of these are cysteine forming 3–4 disulfide bonds. Alpha- and beta- thionins are related to each other. The gamma thionins have a superficially similar structure but are an unrelated class of protein, now called plant defensins.

Recombinant streptavidin-metallothionein chimeric protein having biological recognition specificity

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.
Owner:RGT UNIV OF CALIFORNIA

Prepn of metallothionein

The present invention is one gene engineering process of preparing metallothionein in high yield. The process incldues constituting the expression plasmid or its mutant gene, transferring and fermentation culture in engineering colibacillus, induction and adding metal ion before further culture, collecting thallus; ultrasonic crushing thallus, centrifugation, static adsorption with affinity chromatographic column of glutathione-agarose gel 4B, incision with thrombase on the column, column chromatograhic desalting, gradient salt elution and collecting destination protein. The present inventioncan obtain destination protein with purity over 95 %. Using the said method to prepare 6 mutant proteins of monkey metallothionein I can obtain yield and purity similar to that in wild recombinant protein.
Owner:FUDAN UNIV

Recombination expression carrier and method for soluble expressing human I-type metallothionin

ActiveCN101144093AFermentationHybrid peptidesThioninModifier Genes
The present invention relates to a method for producing a human I type metal sulfur protein (Human metallothionein-I, hMT-I), in particular to the fusion expression by adopting the small molecule ubinquitin-related modifier mature peptide (Small Ubiquitin-related Modifier, SUMO) and hMT-I, and the fusion protein and the ubinquitin-related modifier protease 1 (Ubiquitin-like protease 1) are co-expressed in a prokaryotic organism. In the fermentation and expression process, the ubiquitin associative modifier gene protease 1 can hydrolyze the fusion protein which consists of the ubinquitin-related modifier mature peptide and the hMT-I, to produce the dissoluble hMT-I.
Owner:BIOPHARM RES & DEV CENT JINAN

Recombinant Pseudomonas putida CH01 capable of adsorbing heavy metal cadmium and application thereof

The invention belongs to the soil environment management field in the environmental engineering and relates to the construction and application of a new genetically engineered microorganism capable of restoring the soil polluted by heavy metal cadmium. The preparation method of the genetically engineered microorganism comprises the following steps: monkey metallothionein alpha structure domain tetramer is fused in the ice-nucleating protein N-end of Xanthomona campestris ATCC33913 to be used as a main functional unit for constructing a broad host range expression plasmid; then cadmium-specific promoter is used for expression in Pseudomonas putida X4 which can resist various heavy metals efficiently, thus realizing the surface display of monkey metallothionein alpha structure domain tetramer in the bacterium and obtaining genetically engineered microorganism CH01 which can adsorb heavy metal cadmium efficiently and specifically. The engineered microorganism can remarkably adsorb heavy metal cadmium in the presence of various heavy metals; and the result of the symbiotic water culture with wheat, cowpea and corn shows that the engineered microorganism CH01 can colonize in the rhizosphere of plant and fast adsorb cadmium ions. The invention also discloses a construction method of the genetically engineered microorganism CH01 of which preservation number is CGMCC No.M209320.
Owner:HUAZHONG AGRI UNIV

Enzyme-linked immunosorbent assay method for detecting cadmium-metallothionein in flesh of shell

InactiveCN101900726AThe principle is scientific and reasonableThe technical steps are matureMaterial analysisElectrophoresisCadmium metallothionein
The invention discloses an enzyme-linked immunosorbent assay method for detecting cadmium-metallothionein in the flesh of a shell. The method comprises the following steps of: inducing the shell to synthesize the cadmium-metallothionein by using cadmium chloride, inducing a rabbit to synthesize an antibody by using purified metallothionein, separating and purifying the antibody, using an electrophoretically pure rabbit anti-shell metallothionein antibody as a primary antibody and a goat anti-rabbit enzyme labelled antibody as a secondary antibody, and assaying the cadmium-metallothionein content of the shell by using the indirect competitive enzyme-linked immunosorbent assay method. The method has the advantages of scientific and rational principle and relatively mature technical process and is used for assaying the cadmium-metallothionein content of the flesh of the shell so as to indirectly reflect the cadmium pollution of the flesh of the shell and provide a fast and sensitive detection method for assaying the heavy metal cadmium in the flesh of the shell.
Owner:GUANGDONG OCEAN UNIVERSITY

Streptavidin proteins

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.
Owner:RGT UNIV OF CALIFORNIA

Method for expressing metallothionein of human by using body of vegetable oil

This invention provides a method for expressing human metallothionein (MT) by using plant oil. The method comprises: designing and synthesizing human MT gene according to the preference of plant codon, fusing human MT gene with plant oil protein gene, constructing an expression vector with the fused genes, converting recipient plant by using the expression vector to obtain converted plant, and efficiently expressing human MT by using the seed of the converted plant. Activity experiments show that purified recombinant MT has comparable activity with human origin MT. The method utilizes plant bioreactor to produce MT, thus can avoid possible attack by animal pathogen and toxin in E. coli. The method can largely simplify the separation and purification process of human MT, and lower the production cost, this is advantageous for MT industrialization.
Owner:JILIN AGRICULTURAL UNIV

Cell nucleus DNA (Deoxyribose Nucleic Acid) staining method

InactiveCN102967499AEasy to fixReduce acidolysisPreparing sample for investigation3-deoxyriboseFeulgen stain
The invention discloses a cell nucleus DNA (Deoxyribose Nucleic Acid) staining method. The cell nucleus DNA staining method is characterized by comprising the following steps: 1, fixation: sequentially placing sample slices into a BS fixing solution at the temperature of 30-40 DEG C to fix for 20-30 minutes, and rinsing; 2, hydrolyzation: completely fixing the sample slices, hydrolyzing in 5N hydrochloric acid at the temperature of 30-40 DEG C for 20-30 minutes, and rinsing; 3, staining: after the hydrolyzation is completed, staining in a Thionin staining solution at the temperature of 30-40 DEG C for 30-40 minutes, and rinsing; 4, after the staining is completed, dewatering by adopting fast gradient ethanol, and mounting. In conclusion, compared with other Feulgen staining methods, the cell nucleus DNA cell nucleus DNA has the obvious advantages of practicability, stability, efficiency, cost saving, repeatability, compatibility of automated equipment and the like and can meet the requirement of clinical DNA quantitative cytology on fast diagnosis.
Owner:MOTIC XIAMEN MEDICAL DIAGNOSTICS SYST

Ramie metallothionein gene and recombinant protein and application thereof

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.
Owner:INST OF BAST FIBER CROPS CHINESE ACADEMY OF AGRI SCI

Gene engineering application for elsholtziasplendens metallothionin gene EhMT1

InactiveCN101096672AImprove patienceWith heavy metal binding functionPlant peptidesFermentationNicotiana tabacumThionin
The invention discloses a gene engineering applying method of elsholtzia metal sulphur protein gene EhMT1 in Haizhou, wherein the GenBank entry number of cDNA sequence of EhMT1 is DQ059081 and the coded amino acid sequence is AAY57923; EhMT1 participates the accumulation of Haizhou elsholtzia root; the analysis of mRNA expression is that the Haizhou elsholtzia is induced by excessive copper; EhMT1 can be goal gene leading-in plant, which improves the copper resistance of plant and the accumulation of copper.
Owner:NANJING AGRICULTURAL UNIVERSITY

Method for inducing and extracting metallothionein of the liver of sturgeon

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.
Owner:蓝泽桥

Gene cloning method, expressing method and application of psychrophilic yeast metallothionein

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.
Owner:HARBIN INST OF TECH AT WEIHAI

Recombination expression carrier and method for soluble expressing human I-type metallothionin

ActiveCN101144093BFermentationHybrid peptidesThioninModifier Genes
The present invention relates to a method for producing a human I type metal sulfur protein (Human metallothionein-I, hMT-I), in particular to the fusion expression by adopting the small molecule ubinquitin-related modifier mature peptide (Small Ubiquitin-related Modifier, SUMO) and hMT-I, and the fusion protein and the ubinquitin-related modifier protease 1 (Ubiquitin-like protease 1) are co-expressed in a prokaryotic organism. In the fermentation and expression process, the ubiquitin associative modifier gene protease 1 can hydrolyze the fusion protein which consists of the ubinquitin-related modifier mature peptide and the hMT-I, to produce the dissoluble hMT-I.
Owner:GUANGZHOU BIOPHARMACEUTICAL R&D CENT OF JINAN UNIV CO LTD

Fusion protein of human epidermal growth factor and metallothionein and preparation method and application thereof

The invention provides a fusion protein of a human epidermal growth factor and metallothionein, which comprises a first part and a second part, wherein the first part is the human epidermal growth factor, the second part is the human metallothionein, and the human metallothionein is positioned at the C terminal of the human epidermal growth factor. The invention further provides an application of the fusion protein in the preparation of drugs for treating burns or scalds or wounds or ulcers and the application of the fusion protein in the preparation of skin nursing drugs. Compared with the original human epidermal growth factor and the metallothionein, the fusion protein can increase the stability, reduce the clearance rate, reduce the degradation, bring benefits and convenience to the use of the EGF-MT fusion protein and further reduce the using dosage and the medication frequency. Simultaneously, the formed fusion protein is a bifunctional molecule which has double effects of EGF and MT.
Owner:SHANGHAI SIRUIBAO BIOTECH

Method for detecting peanut metallothionein mRNA expression level by real-time quantitive PCR

The invention discloses a method for a detecting peanut metallothionein mRNA expression level by real-time quantitive PCR, comprising the following steps: 1) connecting metallothionein gene obtained from peanut tissues with vectors, and constructing a recombinant plasmid as a standard substance for real-time quantitive PCR; 2) extracting the recombinant plasmid containing target genes, calculating a molecule copy number, diluting the recombined plasmid into different times to serve as a standard substance template for real-time quantitive PCR reaction; and 3) extracting the total RNA of required tissues, performing reverse transcription on the total RNA to be changed into cDNA serving as a real-time quantitive PCR template, carrying out real-time quantitive PCR to obtain the copy number of an initial template, and detecting and analyzing the mRNA expression level thereof.
Owner:SHANDONG PEANUT RES INST

Yeast strain for displaying metallothionein on cell surface and application of same in heavy metal adsorption

The invention discloses a yeast strain for displaying metallothionein on a cell surface and application of the same in heavy metal adsorption. The yeast strain for displaying metallothionein on the cell surface is obtained by integrating a box gene for displaying metallothionein on the cell surface and a constitutive promoter P<PGK1> gene into the chromosome of a host stain via an integrative vector; and the yeast strain can continuously and stably express metallothionein on the cell surface. The recombinant Saccharomyces cerevisiae strain provided by the invention has good adsorption capability and tolerance to a plurality of heavy metals like Cd<2+>, Cu<2+> and Cr<6+>; e.g., the strain needs 12 h for complete reduction of Cr<6+> with a concentration of 50 mg / L, 6 h faster compared with an original strain, and has a reduction rate 1.5 times of the reduction rate of the original strain; the removal rates of the strain to Cu<2+>, total Cr and Cd<2+> are increased by 13.35%, 10.13% and 19.39% respectively compared with the original strain; so the strain has great application potential in the field of treatment of heavy metals.
Owner:DALIAN UNIV OF TECH

Method for preparing multi-target DNA biosensor using metal ions as markers

InactiveCN103592355ASimple and time-saving preparationQuick responseMaterial electrochemical variablesThioninA-DNA
The invention provides a method for preparing a multi-target DNA (Deoxyribose Nucleic Acid) biosensor using metal ions as markers. According to the invention, the method that the metal ions are used as the markers to construct the DNA biosensor and metallothionein (MT) is used as a bridge to achieve connection of the metal ions and a DNA detection probe sequence is proposed for the first time. Firstly, Zn<2+>, Cd<2+> and Pb<2+> are respectively marked on detection probes of three virus DNA segments by the metallothionein, then the marked detection probes are crossbred with corresponding DNA capture probes fixed on a gold substrate and target DANs in samples to form a sandwich structure, and finally, the metal ions are dissociated by hydrochloric acid and are detected by an anodic-stripping voltammetric method, so that the purpose of detecting the target DNAs is achieved. The sensor prepared by the method can simultaneously detect various target DNAs and has the characteristics of high sensitivity and low limit of detection.
Owner:NORTHWEST NORMAL UNIVERSITY

Human metallothionein-4 fusion protein expression vector

ActiveCN105441472AFacilitate or induce foldingSolving the Inclusion Body ProblemFusions for enhanced expression stability/foldingMetallothioneinsFusion Protein ExpressionThionin
The invention discloses a human metallothionein-4 fusion protein expression vector. The upstream of an expression vector cloning region comprises human heart-type fatty acid-binding protein (hFABP6), and the downstream comprises the human metallothionein-4 (MT-4). The invention also reveals an application method of the expression vector, and the method comprises the following basic steps: construction of an expression bacterial strain, amplification, inducible expression and purification of fusion proteins. The hFABP6-MT4 fusion protein expression vector has the following characteristics (1) high-efficiency expression of MT4 proteins; (2) soluble expression.
Owner:孙袁野

Expression vector and application thereof

The invention discloses an expression vector which comprises at least one regulation unit. Each regulation unit comprises an enhancer and a promoter, wherein the enhancer is an hrx enhancer or functional fragment thereof or sequence variant, and the promoter is an MT promoter or functional fragment thereof or sequence variant, or is an Act5C promoter or functional fragment thereof or sequence variant. The hrx enhancers are hr5 enhancers in Lepidoptera bar viruses AcMNPV, hr3 enhancers in Bombyx mori or hr3 enhancers in AcMNPV. The MT promoters are drosophila cell metallothionein promoters which are induced promoters and induced by metal ions. The Act5C promoters are Actin 5C promoters in drosophila cells. The expression vector can perform efficient expression in drosophila S2 cells. The recombinant protein expression vector with the Lepidoptera bar virus gene hr5 enhancers is built and capable of performing efficient expression in the diptera insect drosophila S2 cells, and overcomes the technical bias that bar viruses are not spread in dipsters due to species isolation in the prior art.
Owner:PROGENBIO CO LTD

Heavy metal toxinicide, its preparation method and application

The present invnetion relates to a biological medicine preparation for relieving heavy metal poisoning. It is a heavy metal antidote using human metallothionein-L (MTL) as main component. The MTL contains the polypeptide of amino acid sequence described by SEQ IP NO2, its nucleotide code sequence and the nucleotide sequence described by SEQ ID NO1 have at least 70% of homology. It can relieve heavy metal and semimetals poisoning, in the course of development it can regulate internal balance of zinc and copper, and make metallic metabolism and detoxification, and can eliminate free radical. There, this invention can be used as auxiliary medicine preparation for curing tumor, and also relates to its production method and application.
Owner:FUDAN UNIV

Metallothionein-like protein gene of Tamarix androssowii Litw.

Get the 3í»cDNA fragment by decreasing treasury, then get the cDNA sequence by cloning with RACE technology, the length of 3 non code area is 262bp, the length of 5 non code area is 150bp, the length of code area is 210bp, containing 69 amino acids, and the molecular weight of code protein is 7.32KD. The metallothionin gene is against heavy metal, drought and salt. The metallothionin is non-enzymic protein which has low molecular mass and high mercapto groups and metals. The said gene is the resistant gene to improve the resistance ability of plant.
Owner:XINJIANG INST OF ECOLOGY & GEOGRAPHY CHINESE ACAD OF SCI

Detecting method of pig metal sulfur protein

The present invention disclosed a detection method of swine metallothionein, relates to detection method of animal metallothionein, it relates to a qualitative and quantitative detection method of swine metallothionein. Said invention uses electrophoretic pure mouse anti-swine metallothionein IgG as first antibody and uses goat anti-mouse enzyme-linked antibody as second antibody, and adopts solid-phase antigen indirect competitive type enzyme-linked immunosorbent assay method to determine the content of swine metallothionein. Said invention also provides the concrete steps of said method.
Owner:HUNAN AGRICULTURAL UNIV

Hyperaccumulator solanum nigrum metallothionein gene (MT-L3) sequence and cloning method thereof

The invention belongs to the field of gene clone in molecular biology, and specifically relates to a hyperaccumulator solanum nigrum metallothionein gene (MT-L3) sequence and a cloning method thereof. The hyperaccumulator solanum nigrum metallothionein gene sequence is type 3 metallothionein gene, and the gene sequence is shown by a nucleotide sequence in SEQ ID NO. 1. The successive cloning and sequencing of the hyperaccumulator solanum nigrum type 3 metallothionein gene helps to establish a base for further analyzing the biological stress resistant capability under the influence of MT-L3, and screening and applying other functions of MT-L3.
Owner:SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI

Plant expression vector utilizing beer yeast MF-alpha to mediate recombinant protein secretion

InactiveCN107653261AGood genetic engineeringGood toolPeptidesFermentationNicotiana tabacumAgricultural science
The invention discloses a plant expression vector utilizing beer yeast MF-alpha to mediate recombinant protein secretion. The plant expression vector contains 35S constitutive promoter, beer yeast MF-alpha gene and exogenous gene. The MF-alpha-exogenous gene in the vector is expressed under control of the 35S constitutive promoter; after the plant expression vector is transferred into wild tobaccoand geranium through agrobacterium mediation, the exogenous gene under control of the 35S constitutive promoter can efficiently express protein coded by the exogenous gene and then secretes the protein out of plants. Experiment results show that yeat MF-alpha signal peptide mediates reporter gene Egfp protein to be secreted out of the plants; when metal sulfur protein MT3 gene of human is used toreplace Egfp gene, it is found that Cd2+ enrichment of transgenic plants is enhanced. Therefore, a genetic engineering operation method can be provided for further building MF-alpha to mediate otherrecombinant proteins to be secreted to the extracellular in the plants.
Owner:KUNMING UNIV OF SCI & TECH

Prepn of metallothionein

The present invention is one gene engineering process of preparing metallothionein in high yield. The process incldues constituting the expression plasmid or its mutant gene, transferring and fermentation culture in engineering colibacillus, induction and adding metal ion before further culture, collecting thallus; ultrasonic crushing thallus, centrifugation, static adsorption with affinity chromatographic column of glutathione-agarose gel 4B, incision with thrombase on the column, column chromatograhic desalting, gradient salt elution and collecting destination protein. The present invention can obtain destination protein with purity over 95 %. Using the said method to prepare 6 mutant proteins of monkey metallothionein I can obtain yield and purity similar to that in wild recombinant protein.
Owner:FUDAN UNIV

Process for extraction preparation of active zinc metallothionein in sea cucumber and abalone

InactiveCN1844141AKeep valid functionsThe extraction process is simple and easyPeptide preparation methodsAnimals/human peptidesSulfurThionin
This invention belonging to metallothionin extraction field is about the extraction procedure of active zinc-sulfur protein from sea cucumber and abalone. The whole procedure involves feedstuff inducing, cleaning the raw material, squashing, filtering, homogenate, crystallization and extraction. With the procedure we reported here, most of sea cucumber and abalone'effective efficiency is remained, the purity of our product is up to 90%, and the procedure is easy to manipulate. We could extract 300g zinc-sulfur protein from 500g dried sea-cucumber or extract 300g active zinc-sulfur protein from 500g alive sea-cucumber.
Owner:YANTAI HAICHENG BIONT

Food-grade lactococcus lactis with human metallothionein-I anchored on surface and preparation method and use thereof

ActiveCN104818227AEliminate the cumbersome steps of separation and purificationReduce lossBacteriaWater contaminantsEscherichia coliThionin
The invention relates to food-grade lactococcus lactis with human metallothionein-I anchored on the surface and a preparation method and a use thereof. The preparation method comprises the following steps of 1, splicing a cA gene and an hMT gene by overlapping PCR, introducing linker between the two genes, and introducing an expression promotion label to an end N to obtain a recombinant cA-hMT gene, 2, connecting the recombinant cA-hMT gene and an expression carrier to obtain a connection product and transferring the connection product into a escherichia coli competent cell, and 3, carrying out inducible expression of the recombinant fusion protein by engineering bacteria obtained by the step 2, carrying out ultrasonication on the obtained engineering bacteria, then carrying out centrifugation and carrying out incubation of food-grade lactococcus lactis by the supernatant so that the fusion protein is anchored on the surface of the food-grade lactococcus lactis without protein purification. The prepared food-grade lactococcus lactis with human metallothionein-I anchored on the surface can enrich heavy metals. The preparation method has the advantages of high efficiency, safety, no toxicity, low cost and no protein purification.
Owner:SOUTH CHINA UNIV OF TECH

A method of differentiating oncogenic changes of the thyroid, a kit to embody the method and the use of metallothionein (mt) for differentiating oncogenic changes of the thyroid

A method of differentiating oncogenic changes of the thyroid encompassing follicular adenoma and follicular cancer, characterised in that in a sample of oncogenic tissue from a patient the expression of the metallothionein (MT) potein is evaluated using immunochistochemical methods using monoclonal anti-MT antibodies, the degree of intensity of the expression of metallothionein is evaluated under a light microscope using the semiquantitative IRS method according to Remmele, wherein if the degree of intensity of the expression of metallothionein is equal to or greater than 8 on the semiquantitative IRS scale then the presence of follicular cancer is indicated.
Owner:弗罗茨瓦夫研究中心有限公司
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