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38 results about "Somatic Cell Hybridization" patented technology

Fusion of 2 somatic cells to form a heterokaryon.

Transgenic rice culture method

The provided specific site can integrates exogenous gene to stable express and affect little to the acceptor. Besides, it also provides a composite gene structure, which can be integrated by rice gene DNA sequence near specific site and DNA sequence of exogenous gene, and be used to culture new rice series by sexual hybridization and somatic hybridization.
Owner:ZHEJIANG UNIV +1

Modified glyphosate-resistant gene and glyphosate-resistant rice cultivation method

The invention belongs to the technical field of plant gene engineering, in particular to a modified glyphosate-resistant gene and a glyphosate-resistant rice cultivation method and comprises modification of glyphosate genes, building of plant expression vectors of the glyphosate-resistant gene and a glyphosate-resistant transgenosis rice cultivation method. Artificially-synthesized modified glyphosate-resistant gene serves as a target gene, the sequence of the gene is as shown in SEQ ID NO: 1, and the target gene is led into rice receptors by an agrobacterium tumefaciens mediated transformation method to obtain high glyphosate-resistant transgenosis rice GT28. the glyphosate-resistant gene of GT28 is recombined by the aid of sexual hybridization and somatic hybridization technique to obtain new rice germplasm, and new glyphosate-resistant varieties (strains) of rice are cultivated.
Owner:HUAZHONG AGRI UNIV

Rapid propagation method for suspension cell culture of Millettia speciosa Champ

The invention discloses a rapid propagation method for suspension cell culture of Millettia speciosa Champ. According to the rapid propagation method, young and tender cotyledons taken from Millettia speciosa Champ pods 50-60 days after flowering are utilized as culture materials, subcultured callus tissue is obtained through inducing culture and subculture, and a stable Millettia speciosa Champ suspension cell culture fluid is obtained through the suspension cell culture. With the adoption of the method, stable Millettia speciosa Champ suspension cells with good quality and high propagation yield can be obtained, and a novel way and method are provided for development and utilization of effective components of the Millettia speciosa Champ and creation of new germplasm resources from the Millettia speciosa Champ with biotechnology methods such as somatic hybridization, genetic transformation, mutant induction and the like.
Owner:GUANGXI BOTANICAL GARDEN OF MEDICINAL PLANTS

Preparation method of catalpa fargesii leaf blade protoplasm

The invention relates to a preparation method of catalpa fargesii leaf blade protoplasts. The catalpa fargesii leaf blade protoplasts are obtained by controlling the ratios and the concentrations of cellulase Onozuka RS and Macerozyme R-10, the concentration of mannitol in enzymatic hydrolysate, enzymolysis temperature, enzymolysis time and other factors. Free protoplasts obtained by the method disclosed by the invention are high in quantity and vitality, and can lay a foundation for researching catalpa fargesii protoplast culture, plant regeneration, genetic transformation, somatic hybridization, protoplast fusion breeding and the like.
Owner:INST OF FORESTRY CHINESE ACAD OF FORESTRY

Cultivating method of new species of anti-late blight and anti-beetle potato

The invention provides a cultivating method of a new species of an anti-late blight and anti-beetle potato, relating to a cultivating method of a new species of the potato with a cell culture technology. The method comprises the following steps of: electrically fusing a protoplast (A) which has anti-late blight gene, anti-beetle gene and anti-oxidation gene, and is separated from the wild species of the potato, with a protoplast (B) of the potato culturing species with wide adaptability to form into a fusion cell with the protoplast A and the protoplast B; generating a callus by means of cell culture, and generating a hybrid test tube plantlet of a regenerated somatic cell by means of induction; and further culturing the tissue culture test tube plantlet, transplanting the cultured tissue culture test tube plantlet into a glasshouse or a land for growing field crops, and generating a test tube first generation somatic cell hybridized potato to obtain the anti-diseases and insect cell hybrid potato, wherein the somatic cell hybridized potato has the good agronomic trait of the potato cultivating species and the anti-late blight and anti-beetle trait. The cultivating method provides a new available gene resource which can be used for improving the quality of the potato and improving the antioxidant activity of the potato cultivating species with the wild species.
Owner:LELING XISEN POTATO IND GROUP

Rubber tree protolast culture plant regeneration method

The invention discloses a rubber tree protolast culture plant regeneration method which comprises the following steps of: establishing an embryonic cell suspension system, separating and purifying protoplast, nursing and culturing the protoplast, carrying out mulitiplication in protoplast regeneration cell clone, inducing somatic embryos and regenerating plants. The rubber tree protolast culture plant regeneration method disclosed by the invention is simple and easy to carry out, the components of a nursing and culture medium employed to nursing and culturing the protoplast are greatly simplified, and the suspension cell systems of the varieties of the rubber tree self are used as the nursing cells, so that the production cost is greatly lowered, the efficiency that the rubber tree protolast is split to form cell clone and the efficiency that the somatic embryos are generated subsequently are effectively improved. A good receptor system is provided for rubber tree variety improvement and quality innovation by utilizing biotechnological breeding modes with significant application prospects, such as protoplast fusion (namely somatic hybridization) and genetic transformation.
Owner:RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI

Cultivating method for transgenic lepidoptera pest resistant rice

The invention belongs to the technical field of rice genetic engineering, and particularly relates to a cultivating method for transgenic lepidoptera pest resistant rice. The method comprises the following steps: using an artificially synthesized novel pesticidal gene cry1C as a target gene, wherein the nucleotide sequence is shown as SEQ ID NO: 1; using a bar gene as a selective marker gene and adopting an agrobacterium mediated genetic transformation method to introduce the target gene into rice excellent restorer minghui 63 to acquire a novel transgenic rice material T1C-19 highly resisting rice lepidoptera pests. Besides, a pest-resistant gene of T19-1 can be recombined into a new rice germplasm through sexual hybridization and somatic hybridization technologies to cultivate a new rice pest resistant product. The invention discloses the verification tests of processes of transformation vecter construction, rice genetic transformation, molecular identification of transformed plants, insertion site sequence analysis, indoor and field pest resistant experiments and the like.
Owner:HUAZHONG AGRI UNIV

Isolated culture method for peony ovules

InactiveCN102511395ASolve the problem of hybrid abortionPlant tissue cultureHorticulture methodsOvaryCulture mediums
The invention relates to an isolated culture method for peony ovules. After peony ovaries which are fertilized for 15 to 90 days are disinfected by mercuric chloride, the ovules are taken out and divided into a 15-to-48-day ovule group and a 49-to-90-day ovule group; the 15-to-48-day ovule group is inoculated into MS culture medium, cultured in the dark for 3 to 15 days, then inoculated into MS culture medium II, cultured in the dark for 10 days and then cultured in light for 7 to 27 days; afterwards, the 15-to-48-day ovule group is inoculated into the MS culture medium II again and cultured in light for 8 to 28 days, so that mature peony seeds I are obtained; and after being first cultured in the dark for 10 days, the 49-to-90-day ovule group is cultured in light for 7 to 27 days, then inoculated in the MS culture medium II again and cultured in light for 8 to 28 days, so that mature peony seeds II are obtained. The isolated culture method for peony ovules can be used in the isolatedresearch on peony embryo development, and solves the problems of the early abortion of the embryos of good-looking peony varieties and the abortion of distant somatic hybrid peonies, and by way of ovule culture, the improved research on peony varieties can be carried out as well.
Owner:HENAN UNIV OF SCI & TECH

Method for transferring Tibetan medicinal effective component into receptor cell using somatic cell cross breeding technology

The process of transferring Tibetan medicinal effective component into receptor cell by means of somatic cell cross breeding technology includes the following main steps: 1) selecting parent material gene types; 2) dissociating the receptor protoplaster; 3) UV irradiating donor protoplaster; 4) fusing the donor protoplaster and the receptor protoplaster and culturing; 5) identifying hybrid callus and regenerated plant; and 6) detecting target characteristic in somatic cell hybrid. The present invention obtains cell line with high medicinal component content and fast growth by means of somatic cell cross breeding technology and may be used in obtaining famous and rare Tibetan medicinal effective component.
Owner:SHANDONG UNIV

Seaweed breeding method based on induction polyploid gametocyte

ActiveCN101911911AIncrease gene contentQuantitative trait genePlant genotype modificationCell hybridizationZoology
The invention discloses a seaweed breeding method based on an induction polyploid gametocyte, which is characterized by comprising the following steps: firstly processing haploid seaweed gametophyte cells by utilizing mitosis hinder agent to lead the chromosome in the cells to be doubled, so as to induce the haploid seaweed gametophyte cells into polyploid seaweed gametophyte cells, and then breeding polyploid seaweed thalluses by utilizing a seaweed gametophyte cell hybridization technique. The method of the invention has the advantages that the seaweed gene content can be improved through polyploid breeding; and the seaweed physiological activity can be strengthened by the increased quantitative character gene, so as to improve economic character thereof. The chromosome doubling method of the invention can be applied for breeding homology polyploids and allopolyploids.
Owner:OCEAN UNIV OF CHINA

Converting diploidy to haploidy for genetic diagnosis

InactiveUS20060166257A1Positively selectableReduce expressionAnimal cellsMicrobiological testing/measurementCancer syndromeMammal
Detection of mutations associated with hereditary diseases is complicated by the diploid nature of mammalian cells. Mutations present in one allele are often masked by the wild-type sequence of the other allele. Individual alleles can be isolated from every chromosome within somatic cell hybrids generated from a single fusion. Nucleic acids from the hybrids can be analyzed for mutations in an unambiguous manner. This approach was used to detect two cancer-causing mutations that had previously defied genetic diagnosis. One of the families studied, Warthin Family G, was the first kindred with a hereditary colon cancer syndrome described in the biomedical literature.
Owner:THE JOHN HOPKINS UNIV SCHOOL OF MEDICINE

Method for establishing hevea brasiliensis suspension cell line

The invention discloses a method for establishing a hevea brasiliensis suspension cell line. The method comprises material selection and disinfection, suspension culture induction process for the callus tissue and a subculture screening culture process. The method for rapidly establishing the hevea brasiliensis suspension cell line, which is provided by the invention, is simple in technological process and relatively short in time consumption, is capable of establishing a stable, uniform, good-dispersity and rapid-growth suspension cell line within only 60-90 days which is reduced by at least a half in comparison with that spent in establishing the hevea brasiliensis suspension cell line by use of a conventional method, greatly lowering the production cost, shortening the experiment period and rapidly providing favorable material sources for technical systems of synchronously inducing regeneration plants from a batch of somatic embryos, or performing protolast culture and somatic hybridization and performing direct conversion on foreign genes by utilizing protoplast as a receptor, and the like.
Owner:RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI

Oncidium protoplast dissociation and culture method

The invention provides an oncidium protoplast dissociation and culture method which comprises the following steps: taking oncidium protocorm-like bodies as starting materials, carrying out callus induction culture and 4-5 times of subculture to obtain vigorously growing calluses with loose structures for dissociation of protoplasts; carrying out cold treatment on calluses at 4 DEG C for 1 d, putting the calluses into a mixed enzyme solution, carrying out shaking table oscillation enzymolysis for 8-10 h, and purifying protoplasts by using an interface method to obtain a large amount of oncidiumprotoplasts with high activity; continuously using the culture medium for solid-liquid double-layer culture, so that the obtained oncidium protoplast cells can normally grow and continuously split to form small cell clusters, the maximum splitting frequency reaches 10.16%, and the maximum plate planting rate reaches 4.03%. The invention provides a scientific basis for cell fusion, somatic cell hybridization, genetic transformation and germplasm innovation by utilizing oncidium protoplast.
Owner:陈俊敏

Plant in-vivo somatic cell hybridization method

ActiveCN114586675ACell culture mediaHybrid cell preparationBiotechnologyCell hybridization
The invention relates to a plant in-vivo somatic cell hybridization method which comprises the following steps: selecting a female organ of a good variety of a plant type expected to be obtained and a somatic cell cluster of a reproduction unit as a hybridization female parent receptor, performing treatment before insemination to remove reproduction isolation, so that the hybridization female parent receptor is easy to fertilize; individuals, organs, tissues, cells, chromosomes or chromosome segments, DNA or DNA segments and genes of animals or plants, microorganisms and other organisms with the characters of the expected variety are selected as male parent donors, a somatic cell hybridization agent is added and stirred evenly, a sperm is obtained, the sperm is used for insemination of a receptor under mediation of the somatic cell hybridization agent, and the variety is obtained. And embryo rescue and seedling culture are carried out after the fertilization embryo is taken back. The bottleneck of distant hybridization is broken through, high-yield and high-quality varieties can be cultivated, and the method has wide application prospects.
Owner:深圳前海觉民科技有限公司

Method for establishing catalpa bungei embryonal cell suspension system

ActiveCN106561452AStrong ability to induce differentiationNot easy to degradeHorticulture methodsPlant tissue culturePolyethylene glycolGermplasm
The invention belongs to the technical field of cell culture, and relates to a catalpa bungei embryonal cell suspension culture method, wherein the method comprises a step of taking embryos of immature fruits of catalpa bungei as explants, and sequentially comprises the steps of inducing embryonal callus, inducing an initial suspension culture product, establishing an embryonal cell suspension system, and regenerating plants; the induction culture medium of the initial suspension culture product is 1 / 2MS; and, the culture solution of the catalpa bungei embryonal cell suspension system is that polyethylene glycol (PEG) 6000 is added based on the 1 / 2MS culture medium. By means of the method disclosed by the invention, the steady catalpa bungei embryonal cell suspension system can be established; furthermore, the plantlet survival rate of embryo regeneration plants is high; emerged seedlings are tidy and normal; and the method disclosed by the invention can provide a steady material source and a platform for catalpa bungei embryo development, somatic hybridization, genetic transformation, and storage and research of plant germplasm resources, and provide important theoretical and experimental basis for industrial breeding of a bioreactor technology based on suspension cell culture.
Owner:CHINA THREE GORGES UNIV

Method for increasing production efficiency of hybrid maize seeds

The invention disclose a method for increasing the production efficiency of hybrid maize seeds. The method includes the steps of S1, culturing maize seeds to obtain maize calluses; S2, separating themaize calluses into a maize protoplast solution; S3, removing the supernate of the maize protoplast solution, and performing precipitation and resuspension; S4, adding maize protoplast into PEG1000, slowly adding 9.5ml of Hanks solution into the test tube 90 seconds later, standing the test tube in a water bath pot, centrifuging and abandoning supernate, adding 1ml of cell culture liquid into thecell precipitates, gently evenly mixing, and culturing in water bath of 37 DEG C for 30 minutes to obtain activated maize protoplast fused cells; S5, using a plant in-vitro tissue culture method to culture the maize protoplast fused cells into regenerated hybrid maize plants. The method has the advantages that maize seeds are bred through a somatic hybridization method, breeding cycle is shortenedgreatly, breeding quality is increased, breeding time is shortened, and the targeting performance of breeding objects is increased.
Owner:安徽富牧通生物科技有限公司

A method for electrofusion of sugarcane protoplasts

InactiveCN109207470AIncrease parental fusion rateImprove one-to-one fusion rateHybrid cell preparationElectrical/wave energy microorganism treatmentPlant LectinsBiotin
The invention belongs to the technical field of plant somatic cell hybridization, in particular to a method for electrofusion of sugarcane protoplasts, comprising the following steps: (1) preparationof parent protoplasts; (2) protoplast inactivation; (3) the addition of protoplast fusion inducer: agglutinating and binding plants to the surface of one parent protoplast and binding biotin to the surface of another parent protoplast; (4) protoplast fusion. The invention utilizes the principle that plant lectin can combine with biotin, plant agglutination and biotin binding to the surface of thetwo parent protoplasts. The two parental protoplasts could be fused by lectin and biotin to improve one-to-one fusion rate. The hybrid progeny possessed the genes of ROC22 sugarcane with high yield, high sugar content and high drought tolerance, and also possessed the genes of GT28 sugarcane with high sugar content and cold tolerance.
Owner:GUANGXI UNIV

A method for sugarcane somatic cell hybridization

ActiveCN109182276AIncrease parental fusion rateImprove one-to-one fusion rateFused cellsCell membranePlant cell
The invention belongs to the technical field of plant somatic cell hybridization, in particular to a method for sugarcane somatic cell hybridization, comprising the following steps: (1) preparation ofparent protoplast; (2) protoplast inactivation; (3) the addition of protoplast fusion inducer: one parent protoplast was cultured in silicate solution and mixed with another parent protoplast to obtain mixed protoplast; (4) fusion of protoplasts: electrofusion of the mixed protoplasts obtained in step (3). The invention utilizes the principle that silicon has high affinity with glycolipid on thesurface of plant cell membrane, The two parent protoplasts fused under the induction of silicon to improve the one-to-one fusion rate. The hybrid progenies had the genes of ROC22 sugarcane with high yield, high sugar content and high drought tolerance, and had the genes of GT28 sugarcane with high sugar content and cold tolerance.
Owner:GUANGXI UNIV

Method for cultivating insect-resistant transgenic rice T1c-19

The invention belongs to the technical field of genetic engineering for rice, and particularly relates to a method for cultivating insect-resistant transgenic rice T1c-19. The method includes transferring target genes into excellent rice restorer lines minghui 63 by the aid of synthesized novel insecticidal genes cry1C, bar genes and agrobacterium tumefaciens mediated genetic transformation processes to obtain novel transgenic rice materials T1C-19 with high rice Lepidoptera insect resistance. The novel insecticidal genes cry1C are shown as SEQ ID NO:1 and are used as the target genes, and the bar genes are used as selectable marker genes for transferring the target genes into the excellent rice restorer lines minghui 63. The method has the advantages that the insect-resistant genes of T19-1 further can be recombined into novel rice germ plasma by the aid of sexual hybridization and somatic hybridization technologies, flanking sequences of insertion fragments of exogenous genes of transgenic plants of the rice are 5'-end flanking sequences shown as SEQ ID NO:3 and or 3'-end flanking sequences shown as SEQ ID NO:5, and the method for completely cultivating novel insect-resistant varieties (strains) of rice is provided; the T1C-19 is a novel genetic resource.
Owner:HUAZHONG AGRI UNIV

Application of thinopyron elongatum in formulating low-celiac syndrome antigen encoding DNA wheat

InactiveCN101173267ASolve food problemsFermentationGenetic engineeringEpitopeTriticeae
The invention discloses an application that low celiac disease epitope coding DNA wheat can be created using couch grass with long ear. The application screens out distant grass of couch grass with long ear for low celiac disease epitope coding DNA wheat using a homologous sequencing; gene group or DNA of distant grass part is transferred into wheat using somatic cell hybridization technology before low celiac disease epitope coding DNA wheat somatic hybrid is screened out using the homologous sequencing. The somatic hybrid can be used for breeding work of low celiac disease epitope wheat and lay the foundation for food problem during therapy period for celiac disease patients.
Owner:SHANDONG UNIV

Method for preparing high-quality rehmannia protoplast

The invention discloses a method for preparing high-quality rehmannia protoplast, which comprises the following steps: (1) rehmannia leaf pretreatment: taking rehmannia seedling leaves, cutting the rehmannia seedling leaves into thin strips, and soaking the thin strips in a buffer solution in a dark place; (2) enzymolysis: transferring the pretreated rehmannia leaves into an enzymatic hydrolysate, and oscillating in a dark place for enzymolysis; (3) protoplast separation; (4) purifying the protoplast: resuspending the separated protoplast by using a buffer solution; adding the resuspended liquid to the upper layer of the sucrose salt solution, and centrifuging; after centrifugation, the middle layer is a protoplast band. The method is simple, the prepared protoplast is regular in shape, complete in structure, high in living protoplast content and good in quality, and a foundation is laid for research in the fields of heredity and molecular biology, such as genetic transformation, rapid propagation, somatic cell hybridization and breeding of good varieties by using the rehmannia protoplast as an experimental material.
Owner:HENAN UNIVERSITY OF TECHNOLOGY

Method for transferring Tibetan medicinal effective component into receptor cell using somatic cell cross breeding technology

The process of transferring Tibetan medicinal effective component into receptor cell by means of somatic cell cross breeding technology includes the following main steps: 1) selecting parent material gene types; 2) dissociating the receptor protoplaster; 3) UV irradiating donor protoplaster; 4) fusing the donor protoplaster and the receptor protoplaster and culturing; 5) identifying hybrid callus and regenerated plant; and 6) detecting target characteristic in somatic cell hybrid. The present invention obtains cell line with high medicinal component content and fast growth by means of somatic cell cross breeding technology and may be used in obtaining famous and rare Tibetan medicinal effective component.
Owner:SHANDONG UNIV

Method for preparing amaranthus protoplast and application of amaranthus protoplast

PendingCN113897328APromote genetic breedingPromote the development of basic researchFused cellsFluorescence/phosphorescenceBiotechnologySubcellular localization
The invention provides a method for preparing amaranthus protoplast. The method is characterized in that tender true leaves of amaranthus sterile seedlings are used as materials, filter residues are extruded during filtration after enzymolysis, a large number of amaranthus protoplast with normal form, few impurities and no damage is prepared, the experimental requirements of subsequent conversion, subcellular localization and the like can be met, and important technical support can be provided for fundamental researches and application researches such as dual-fluorescence complementation, somatic cell hybridization and genetic transformation, so that the development of genetic breeding and fundamental researches of the edible amaranthus is promoted.
Owner:BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES

A kind of establishment method of catalpa catapulia embryogenic cell suspension line

ActiveCN106561452BStrong ability to induce differentiationNot easy to degradeHorticulture methodsPlant tissue cultureGermplasmPolyethylene glycol
The invention belongs to the technical field of cell culture, and relates to a catalpa bungei embryonal cell suspension culture method, wherein the method comprises a step of taking embryos of immature fruits of catalpa bungei as explants, and sequentially comprises the steps of inducing embryonal callus, inducing an initial suspension culture product, establishing an embryonal cell suspension system, and regenerating plants; the induction culture medium of the initial suspension culture product is 1 / 2MS; and, the culture solution of the catalpa bungei embryonal cell suspension system is that polyethylene glycol (PEG) 6000 is added based on the 1 / 2MS culture medium. By means of the method disclosed by the invention, the steady catalpa bungei embryonal cell suspension system can be established; furthermore, the plantlet survival rate of embryo regeneration plants is high; emerged seedlings are tidy and normal; and the method disclosed by the invention can provide a steady material source and a platform for catalpa bungei embryo development, somatic hybridization, genetic transformation, and storage and research of plant germplasm resources, and provide important theoretical and experimental basis for industrial breeding of a bioreactor technology based on suspension cell culture.
Owner:CHINA THREE GORGES UNIV

A kind of method of sugarcane somatic cell hybridization

ActiveCN109182276BIncrease parental fusion rateImprove one-to-one fusion rateFused cellsBiotechnologyPlant cell
The invention belongs to the technical field of plant somatic cell hybridization, in particular to a method for sugarcane somatic cell hybridization, comprising the following steps: (1) preparation of parental protoplasts; (2) passivation of protoplasts; (3) protoplast fusion induction The addition of thing: wherein a kind of parent protoplast is cultured with silicate aqueous solution, mixes with another kind of parent protoplast equal volume to obtain mixed protoplast; (4) fusion of protoplast: the step (3) obtains Mix protoplasts for electrofusion. The invention utilizes the principle that silicon has a high affinity with glycolipids on the surface of plant cell membranes. The two parental protoplasts are fused under the induction of silicon to increase the one-to-one fusion rate. The hybrid offspring also has high yield and high sugar in ROC22 sugarcane , high drought tolerance genes, and high sugar cold tolerance genes of GT28 sugarcane.
Owner:GUANGXI UNIV
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