64results about How to "Increase genetic diversity" patented technology

The invention relates to gene engineering and antibody preparation and specifically discloses a preparation method and application of a high-throughput fully-humanized antibody. The preparation methodcomprises the following steps: separating peripheral blood monouclear cells; separating single cells of plasma cells or antigen-specificity memory B blymphocytes; amplifying the heavy-chain and light-chain gene variable regions of single B cell antibody by use of a primer provided by the inventor; establishing an expression system containing antibody heavy-chain and light-chain genes by use of alinear expression system containing a heavy-chain fragment or light-chain fragment constant area; finally, separating and purifying a fully humanized monoclonal antibody. By adopting the primer combination provided by the invention to amplify the genes in the antibody heavy-chain and light-chain variable areas, natural pairing of the light-chain and heavy-chain variable areas can be maintained, and the preparation method has the advantages of high gene diversity, high titer, full humanization, high antibody affinity, strong specificity, no heterologous serum reaction, no risk of propagating other infectious diseases, etc.

The invention relates to gene engineering and antibody preparation and specifically discloses a preparation method and application of a high-throughput fully-humanized antibody. The preparation methodcomprises the following steps: separating peripheral blood monouclear cells; separating single cells of plasma cells or antigen-specificity memory B blymphocytes; amplifying the heavy-chain and light-chain gene variable regions of single B cell antibody by use of a primer provided by the inventor; establishing an expression system containing antibody heavy-chain and light-chain genes by use of alinear expression system containing a heavy-chain fragment or light-chain fragment constant area; finally, separating and purifying a fully humanized monoclonal antibody. By adopting the primer combination provided by the invention to amplify the genes in the antibody heavy-chain and light-chain variable areas, natural pairing of the light-chain and heavy-chain variable areas can be maintained, and the preparation method has the advantages of high gene diversity, high titer, full humanization, high antibody affinity, strong specificity, no heterologous serum reaction, no risk of propagating other infectious diseases, etc.

The invention discloses a breeding method of a high-oleic-acid dormancy peanut variety. The breeding method of the high-oleic-acid dormancy peanut variety comprises the following steps: hybridizing parents, namely taking dragonhead No.2 as a female parent and taking a high-oleic-acid dormancy peanut variety system P76 as a male parent; screening high oleic acid by using pressure, namely harvesting single plants of F2-F3 hybridized generations of seeds, carrying out detection on the content of the nondestructive oleic acid in all the seeds of the single plates commonly and carrying out two generations of screening; screening high dormancy by using pressure; testing the dormancy and testing the content of the nondestructive oleic acid; and obtaining the high-oleic-acid dormancy peanut variety. The bred peanut variety is high in content of oleic acid and high in dormancy; the problem that sprouts grow on the plants when harvesting the peanuts can be effectively solved.

The invention discloses a method for quickly establishing an improved breeding system of YY super-male pelteobagrus fulvidraco. The method mainly includes the first step of screening pelteobagrus fulvidraco wild male parents, the second step of carrying out wild parent germplasm resource evaluation on the pelteobagrus fulvidraco, the third step of establishing YY female and YY male basic breeding population through mating with existing YY males, the fourth step of carrying out breeding on the YY female and YY male basic breeding population, the fifth step of carrying out test mating verification on the YY females and the YY males, the sixth step of carrying out quick population expansion on the improved breeding system of the YY female super-male pelteobagrus fulvidraco and establishing the improved breeding system of the YY female super-male pelteobagrus fulvidraco, and the last step of repeating the sixth step, so the improved breeding system can be established. According to the method, genetic diversities of the super-male pelteobagrus fulvidraco are increased, the rate of fertilization, and the incubation rate and the survival rate are improved; meanwhile, the breeding time is shortened, batch breeding can be achieved, the YY female super-male pelteobagrus fulvidraco and XY male fishes with improved characters can be produced in a large-scale mode, the yield can be improved greatly, and economic benefits can be increased.

The invention provides an SNP (single nucleotide polymorphism) markers based on a KASP (kompetitive allele-specific polymerase chain reaction) and application of the SNP markers and belongs to the technical field of molecular biology, germplasm resources and molecule breeding techniques. The SNP markers comprise 48 markers such as CigarSNP01-1, CigarSNP01-2 and CigarSNP02-1. By adopting the SNP markers provided by the invention, cigar SNP fingerprint spectrum construction can be completed, conventional cigar resources can be systematically identified, repeatedly collected resources can be alsorejected, and intellectual property protection mechanisms of cigar varieties can be constructed. The SNP markers provided by the invention are adopted to screen and identify collected cigar resourcesand examine and approve newly cultured cigar varieties, and are applied to cigar breeding operation.

The invention discloses a Honglian type new gene restoring method which utilizes molecule marker to detect the three-line hybrid rice, comprising the steps as follows: first, the design of the specific marker primer; second, the trace extraction of DNA; third, the construction of standard PCR expansion system; forth, the detection analysis of the expanded molecule marker; fifth, the determination of new restoring gene. The Honglian type new gene restoring method is based on the molecule marker of restoring gene co-segregation; the rice germplasm material is screened fast, accurately and reliably in laboratory. The Honglian type new gene restoring method has the advantages of simple method, convenient operation, strong purpose and pertinence for breeding of new restoring line, and increased breeding efficiency of new restoring line.

The invention discloses a method for keeping the genetic diversity of Jian carps. The method comprises the steps of individual tagging, template creating, genetic typing, family building, breeding file establishing and parent updating. Compared with the prior art, inbreeding of the Jian carps can be effectively avoided, the genetic diversity of Jian carp groups is kept and increased, and genetical characterization decline of the Jian carps is slowed down; in addition, breeding is assisted through molecular markers, a breeding file is established, genetic distances of the groups are analyzed by combining all parameters in the file, and therefore dominant groups are selectively reserved, and descendants excellent in genetic characterization are obtained through mating.

The invention discloses a paternity test method of common suckers. The method comprises the following steps of: (1) extracting DNA from common sucker individuals; (2) screening microsatellite markers of the common suckers; (3) carrying out synthesizing and PCR (polymerase chain reaction) amplification on the microsatellite fluorescence markers of the common suckers; and (4) creating paternity test technology of the common suckers. A paternity test kit of the common suckers comprises 10mM / L of 10*PCR Buffer, 25mmol / L of MgCl2, 10mM / L of dNTPs, 10mM / L of 17 microsatellite markers (F.R), 5U / microliter of rTaq Enzyme and ddH2O. The paternity test method and the kit of common suckers disclosed by the invention can be used for realizing artificial reproduction of common sucker farms, providing reference for creating a common sucker family tree, guiding the artificial fecundation and release of the common suckers and improving the reproduction survival rate of the common suckers.

The invention discloses a herbicide-resistance-based rape hybrid seed producing method and applications thereof, and relates to the field of plant seed production. The method includes (1) introducing a wide-spectrum type herbicide-resistant gene into a rape male parent line to obtain a herbicide-resistant rape male parent line that is an imidazolidinone-resistant line 12WH381 or a glufosinate-resistant line Z7B10, (2) pollinating a female parent that is sterile after a chemical androcidal action by utilizing pollen of the herbicide-resistant rape male parent line, and harvesting hybrid seeds from male parent plants, and (3) coating the hybrid seeds with a herbicide, or spraying a corresponding herbicide to the hybrid seeds when the hybrid seeds are planted to remove false hybrid seeds at the same time of removing weeds so as to obtain high-purity rape hybrid seeds or group. The applications include (1) hybrid seed production for rape, (2) improvement on rape three-line seed production methods at present, (3) improvement on rape nucleus-sterility two-line seed production methods at present and (4) improvement on rape chemical androcidal seed production systems at present.

The invention relates to a microsatellite molecular marker-specific primer of litopenaeus vannamei, which can be named as LavTF001, LavTF003, LavTF005, LavTF006, LavTF007, LavTF015, LavTF018, LavTF022, LavTF025, LavTF028 and LavTF029, and comprises 22 specific primer sequences. The microsatellite marker is stable and reliable, and can be used for genetic diversity analysis, germplasm identification and evaluation, functional gene mapping and molecular breeding research of litopenaeus vannamei population.

The invention provides a breeding method for inducing intersexual pelteobagrus fulvidraco to generate a super-male fish synthetic line and an all-female synthetic line. The breeding method comprises the steps of induction of gamete XY type intersexual fish and gamete XX type intersexual fish and criterions of gamete types, a test cross screening method that backcross of the XY type intersexual fish is utilized to generate a candidate super-male fish population and super-male fish, and a technology that backcross of the gamete XX type intersexual fish is utilized to generate the all-female synthetic line. According to the technology, for multiple geographical populations introduced according to the technical requirements, the super-male fish synthetic line and the all-female synthetic lineare constructed respectively, and a diallel crossing test is used for screening an all-female pelteobagrus fulvidraco production combination with hybrid vigors. The breeding method for inducing the intersexual pelteobagrus fulvidraco to generate the super-male fish synthetic line and the all-female synthetic line has the advantages that an electronic digital chip tag labeling management method isestablished in order to prevent population mixing and ensure the accuracy of test cross identification, the induced intersexual fish does not need genetic identification, a screened super-male familyhas high male rate, the requirements for instrument equipment are simple, the technology is easy to operate, and the breeding method is also suitable for breeding of all-male fish and all-female fishof other male gamete alien fish.

The invention provides a method for developing polymorphic microsatellite molecular markers on the basis of multi-sample high-throughput sequencing. The method comprises the steps as follows: firstly,high-throughput sequencing data of multiple sample genomes or transcriptomes of to-be-tested species are acquired, potential satellite sits capable of being amplified are screened, microsatellite sites with primer pairs appearing only once in all sequencing reads of the potential satellite sits capable of being amplified in single sample are screened, and markers which appear in multiple samplesand have the allele number larger than 2 in all the samples are screened; finally, those markers appearing in reference genomes or transcriptomes only once are screened by establishing a reference sequence database, and those repeated markers amplifying the same fragments are eliminated. The polymorphic microsatellite markers can be effectively screened by use of high-throughput sequencing data, non-specific and repeated markers can be removed, the length of PCR amplified products of corresponding microsatellite sites can be predicted, and the efficiency of development of microsatellite molecular markers is significantly improved.

The invention belongs to the technical field of paddy rice breeding and discloses a paddy rice breeding method. According to the paddy rice breeding method, an imported foreign material is hybridizedwith a local variety; after different generations of separation and selection are carried out, properties are listed as follows: a combination (1): IR71121-35-1-1-1-2 / V13 hybridized F4 has the plant height of 80 cm, the quantity of total grains per ear is 246.7 and the fruiting rate is 73.4 percent; a combination (2): IR64446-2 / Dianjingyou No. 5 hybridized F5 has the plant height of 90 cm, the quantity of total grains per ear is 189.0 and the fruiting rate is 93.5 percent; a combination (3): Taebongbyeo / Undong 31 / / Yujing No. 8 hybridized F3 has the plant height of 90 cm, the quantity of totalgrains per ear is 203.0 and the fruiting rate is 91.0 percent. The paddy rice breeding method provides abundant paddy rice breeding resources for scientific researches and variety improvement of riceculture; the intra-regional and inter-regional genetic diversity is further expanded and a new leap of the production of paddy rice is realized.

The invention relates to a high-oleic acid peanut mutant gene AhFAD2B-814 and application thereof and belongs to the field of plant molecular breeding. A nucleotide sequence of the mutant gene AhFAD2B-814 is shown as SEQ ID NO.: 1 and consists of 1140 bases. The high-oleic acid peanut mutant gene AhFAD2B-814 and the application thereof provided by the invention have the benefits that a mutant nucleotide sequence of a peanut high-oleic acid mutant oleic acid dehydrogenase gene can be introduced into other peanut varieties or strains with common oleic acid contents by applying conventional breeding methods such as hybridization, backcrossing and the like, the oleic acid content of a target variety or strain introduced with the mutant nucleotide sequence of the gene AhFAD2B-814 is improved, the genetic diversity of a high-oleic acid peanut variety (strain) is significantly increased, and the genetic basis of a high-oleic acid peanut is widened; during high-oleic acid peanut breeding, by selecting a peanut material carrying an AhFAD2A mutant genotype as a parent, the selection on high-oleic acid characters of a breeding offspring is more purposeful, and the occurrence frequency of target traits is improved.

The invention discloses a microsatellite marker relevant with tachysurus fulvidraco growth characteristics and detection and application thereof. That is, four microsatellite loci are found on a tachysurus fulvidraco GH sequence, the microsatellite loci are amplified by screening specific primers, relevance between the microsatellite loci and growth traits is analyzed, and dominant genotypes favorable for the growth traits are found. Results indicate that three loci are remarkably or extremely remarkably relevant with the growth traits, a locus GA is remarkably relevant with body weight, body length, standing height and body thickness and is extremely remarkably relevant with total length and depth of caudal peduncle, wherein CD is a dominant genotype; a locus TCTT is remarkably relevant with head length and depth of caudal peduncle, wherein BD is a dominant genotype; and a locus AC is remarkably relevant with body thickness and head length, wherein DD is a dominant genotype. The four microsatellite loci obtained by primer amplification have high genetic diversity and are remarkably or extremely remarkably relevant with the growth traits. The microsatellite marker is of great significance for molecular breeding practice.

The invention relates to a method for matching and breeding jian carps. The method comprises the following steps of: sampling blood or fin rays of a jian carp breeding population to extract genome DNA; detecting the genotype of SNPs (Single Nucleotide Polymorphisms) by a PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism) method constructed by taking the extracted genomeDNA as a template according six SNP sites related to jian carp weight increment; carrying out genotype determination by using a sepharose gel electrophoresis enzyme slitting strip; selecting individuals containing more than four genotypes with rapid weight increment as candidate parents; carrying out digital treatment on the candidate parents by using the PCR-RFLP sepharose gel electrophoresis enzyme slitting strip; analyzing by using genetic analysis software to obtain genetic distances between every two individuals; constructing an evolutionary tree; and selecting an individual which has a larger genetic distance and better gonad development and can enrich more than four genotypes with rapid weight increment. The method is helpful to improving the production performance of jian carps, avoiding inbreeding and accelerating the breeding process of jian carps.

The invention discloses a Rhinogobio ventralis Sauvage et Da bry paternity testing method based on micro-satellite fluorescence multiplex PCR. The Rhinogobio ventralis Sauvage et Da bry paternity testing method comprises: 1, extracting the genomic DNA of a to-be-tested sample; 2, carrying out a multiplex PCR amplification reaction by using a primer combination, and carrying out allele typing by using a sequencer; and 3, detecting the correlation between a to-be-tested subject and a parental genotype through likelihood ratio, and determining the paternity. Compared to the traditional PCR method, the method of the present invention has the following advantages that the detected microsatellite loci are increased by about 3 times so as to substantially improve the efficiency and the speed of the paternity testing, the detection cost is only about 1 / 3 of the original detection cost, the allele size interpretation error and other problems in the typing using the acrylamide gel electrophoresis are overcome, the accuracy of the genotype data is improved, and the analysis results show that 100% of the to-be-tested individuals can correctly find the parents.

The invention discloses a method for long-term storage of berberis heteropoda seeds. The method comprises the following steps that the seeds are sterilized, disinfected and aired, and the dried berberis heteropoda seeds are placed under a 2-5 DEG C condition and soaked in a mixed solution comprising 15% of ethanediol, 10% of dimethyl sulfoxide (MSO) and 10% of cane sugar for 45-60 minutes; prefrozen berberis heteropoda seeds are placed into an absorbent gauze bag, and the berberis heteropoda seeds and the gauze bag are placed into a dryer with the lower layer being made of silica gel for drying; and the dried seeds are uniformly smeared with ceresin, packaged with a tinfoil bag airtightly, placed into a low-temperature environment with the temperature being 25-40 DEG C to be stored for 30-60 minutes, and then packed into a sealed bag which is filled with liquid nitrogen and has the temperature of -196 DEG C for cryopreservation. By adoption of the method, the activity of the seeds can be preserved in a long term, the rate of emergence of the seeds is guaranteed and increased, and the seedling establishment capability is improved.

The invention relates to a preparation method of a pig-source single-chain genetic engineering antibody against foot-and-mouth disease viruses, and belongs to the technical field of preparation of antibodies. PBMCs are isolated from an immune pig body with a foot-and-mouth disease virus antigen, an FMDV 146 antigen is marked with a dyestuff, the PBMCs are dyed, and a single B cell combined with FMDV is sorted by virtue of a multicolor flow cytometer; combined with single-cell antibody gene sequencing, VH and VL genes are obtained respectively, and through a flexible linker and CHO-S expressionengineering antibody technology, the FMDV specific pig-source single-chain genetic engineering antibody is obtained through expression. The antibody obtained by the preparation method has the advantages of good genetic diversity, high efficiency, whole-host (pig) source, few needed cell quantity and the like.

The invention relates to a gene QDTY 2.9IR66897B capable of obviously increasing rice reproductive-stage drought tolerance and a molecular marker method thereof, belonging to the fields of rice high-yield stress-tolerant breeding and molecular genetics. By utilizing Jijing 88 and 3 donor-derived breeding population selection, a novel segregation distortion locating process suitable for breeding population selection is adopted to locate drought-tolerance-related QTL (quantitative trait loci); and random population selection is utilized to verify the authenticity of the drought-tolerant QTL and the reliability of the close linkage marker. When being applied to auxiliary selective breeding and polymerization breeding of drought-tolerant rice, the gene can effectively overcome the defects in the past spontaneous mutant genes, can obviously lower the negative influence on the auxiliary breeding due to the development of the initial locating result, and can carry out genotype selection on the lower-generation breeding populations in the seedling stage to obtain the drought-tolerance individuals, thereby facilitating the timely hybrid transformation, omitting the drought resistance identification process in the adult-plant stage, enhancing the breeding efficiency and accelerating the breeding progress.

The invention belongs to the technical field of biology, and particularly relates to a full-swine-origin African swine fever virus monoclonal antibody as well as a preparation method and application thereof. The monoclonal antibody comprises heavy chain constant regions, heavy chain variable regions, light chain constant regions and light chain variable regions, the amino acid sequences of the heavy chain variable regions are as shown in SEQ ID NO: 1 or SEQ ID NO: 5, and the amino acid sequences of the light chain variable regions are as shown in SEQ ID NO: 2 or SEQ ID NO: 6. The full-swine-origin monoclonal antibody specifically aiming at the African swine fever virus retains natural pairing of light chain and heavy chain variable regions, has the advantages of good gene diversity, full swine origin, good antibody affinity, strong specificity and the like, and can be efficiently expressed in a eukaryotic expression system. According to the invention, a novel raw material is provided for developing a diagnostic kit for African swine fever virus and a new technical support is also provided for developing a preventive and therapeutic drug of a full-swine-origin monoclonal antibody.

The invention belongs to the technical field of biology, and particularly relates to a wild population restoration method of an endangered rheum plant - rheum nobile which has the ornamental value andmedical value. The method comprises the following steps: performing the manual cross-pollination to increase the seed yield and overcome the limitation of seed insufficiency caused by the biting andeating of a large number of seeds by insects under a natural pollination condition, scientifically detecting a development situation of the seeds by using a digital X-ray sample imaging system to improve the quality of the to-be-planted seeds, selecting or creating an appropriate seeding land to increase the emergence rate, and scientifically managing at the seedling period to increase the survival rate of seedlings. The invention relates to the rheum nobile wild population restoration technology which is simple in operation, convenient and easy, convenient in management and capable of savingworkers and manpower, thereby providing technical support for the protection and sustainable utilization of Tibet medicine plant resources.

The invention discloses rice uracil DNA (deoxyribonucleic acid) glycosidase and application thereof in gene editing induction of plant single base diversity. The rice uracil DNA glycosidase is a protein composed of an amino acid sequence as shown in SEQ ID NO. 2; or a protein which is derived from the amino acid sequence shown in SEQ ID NO.2 and keeps the protein function shown in SEQ ID NO.2 through substitution and / or deletion and / or addition of one or more amino acid residues of the amino acid sequence shown in SEQ ID NO.2. According to the application provided by the invention, a biological material containing the rice uracil DNA glycosidase UNG is constructed into a plant cytosine single-base editing vector OsCGBE containing the OsUNG sequence, the expression vector is introduced into a plant, a C base on DNA is induced to be mutated into T or G or A, plant single-base diversity is efficiently created, and the application range of a single-base editor is expanded.

A system for determining traits for an individual mammal includes a first database with genomic data of known traits as well as identified correlations including details of the trait, mutation, disease risk and / or affected breeds of a mammal species, and a second database based on a gene test made for the individual mammal, the database including genotyping data of the individual mammal to be analyzed. A plurality of markers is determined for regions in the mammal's genome so that at least first portion of the markers relates to correlations in various known gene based traits, and at least second portion of the markers used in determining differs from the first portion of the markers. Then the genotyping data of the individual mammal's genome corresponding to the first portion of the markers is compared in order to make correlations and thereby determine a probability or risk or severity of the traits.

The invention specifically relates to a winter wheat region wheat synthesizing method with high genetic diversity and a high embryo formation and seedling formation rate and solves the problems that existing synthesized wheat is not suitable for winter wheat regions, genetic information is limited, and the embryo formation and seedling formation rate is low. The winter wheat region wheat synthesizing method comprises the steps: a, choosing durum wheat and aegilops tauschii to be sowed; b, pollinating pollen of the aegilops tauschii to the durum wheat; c, performing cultivation and sterilization; d, inoculating young embryo of sterilized kernels to a culture medium II to be cultured; e, performing vernalization culture when seedlings are achieved; f, transplanting the seedlings to a land for growing field crops; g, performing soaking treatment through a colchicine solution with the concentration as 1% when the seedlings grow out tillers and then continuing performing field culture and management until the wheat is harvested. The synthesizing method disclosed by the invention is suitable for synthesizing and breeding a wheat novel material of the winter wheat region of northern China.

The invention discloses a method for hybridizing soybeans in tropical regions. The method comprises the following steps: 1, firstly, sowing a male parent, then sowing a female parent, and carrying light treatment on the male parent and the female parent for realizing the flower synchronization of the male and female parents after soybean germination according to the following method: carrying out light supplementing treatment on the male parent, wherein the daily lighting time is 17-18 hours; arranging the female parent in a natural light condition without light supplementing treatment, wherein the daily lighting time is 11-12 hours; and 2, in the flowering periods of the male and female parents, removing a corolla of a flower on a female parent plant but maintaining stamen, pollinating the pollen of the male parent on a stigma of the female parent, and thus obtaining a hybrid seed on the female parent. The method is utilized to hybridize the soybeans in tropical regions, the hybridizing efficiency can be increased, processed are reduced, and the cost is lowered; and the method has important meanings in improving the genetic diversity of improved soybean varieties and accelerating a soybean seed breeding process.

The invention discloses a multiplex amplification detection system and method including 13 rapidly-mutating Y-STR loci and application. The multiplex amplification detection system includes the 13 loci and fluorescent labeling dyes corresponding to primers of the 13 loci. The invention develops a set of multiplex amplification detection system including the 13 rapidly-mutating Y-STR loci, and a five-color fluorescent technique (blue FAM, green HEX, yellow TAMRA, red ROX and orange) is selected to perform multiplex amplification, so that good amplification: a peak pattern is sharp, no blended bands are generated and the balance is good, of a DNA sample can be realized. The system can be applied to refined identification in male family screening, and provides a technical support for investigation and case handling.