127 results about "Nuclear gene" patented technology

The current invention provides the promoter of the Zea mays nuclear gene encoding chloroplast-localized fructose-1,6-bisphosphate (F16BP) aldolase. Compositions comprising this sequence are described, as are plants transformed with such compositions. Further provided are methods for the expression of transgenes in plants comprising the use of these sequences. The methods of the invention include the direct creation of transgenic plants with the chloroplastic F16BP aldolase promoter by genetic transformation, as well as by plant breeding methods. The sequences of the invention represent a valuable new tool for the creation of transgenic plants, preferably having one or more added beneficial characteristics.

Gene targeting allows the deletion (knock out), the repair (rescuing) and the modification (gene mutation) of a selected gene and the functional analysis of any gene of interest. Targeting of nuclear genes has been a very inefficient process in most eukaryotes including plants and animals due to the dominance of illegitimate integration of the applied DNA into non-homologous regions of the genome. The present invention provides a method for increasing the ratio of homologous to non-homologous recombination of a polynucleotide into a host cell's DNA by suppressing non-homologous recombination. Surprisingly, the number of non-homologous recombination events can be reduced if the polynucleotide is applied as a purified single-stranded DNA, preferably coated with a single strand binding protein.

The invention provides a molecular marker for identifying amur sturgeon germplasm and a method for identifying amur sturgeon germplasm by virtue of the molecular marker. Amur sturgeon or a finished product thereof can be directly identified through sequence alignment on the basis of an interspecific sequence difference of an fzd8 gene segment obtained from PCR amplification of a specific primer between the amur sturgeon and other sturgeons. The molecular marker disclosed by the invention, which is used for analyzing and identifying an SNP site on the fzd8 nuclear gene segment screened from acipenseridae fishes, has the advantages of being accurate in experiment, reliable and rapid in identification, convenient and simple in operation, and the like.

The invention relates to an ARMS fluorescent quantitative PCR-based gene mutation kit and a method thereof. Compared with present gene polymorphism detection kits and methods thereof, the enhanced ARMS fluorescent quantitative PCR-based gene polymorphism mutation kit and the method thereof have the advantages of strong specificity, high sensitivity, low cost, highly reliable result, low background, easy determination of the result, ascendant operation and use, fast and convenient fluorescent quantitative PCR, closed tube detection in the whole course, pollution reduction, and reduction of the unnecessary cost and workload. The fluorescent quantitative PCR can realize the characterization of the amplification efficiency by using a CT value, so the method can realize the detection of the nuclear gene polymorphism heterozygote genotype conditions, and makes the result accurately and reliably determined.

The invention discloses a probe fluorescent quantitation polymerase chain reaction (PCR) method for rapidly detecting pork or chicken compositions in food added with internal amplification control. The method includes designing a primer and a probe respectively based on an animal nuclear gene; and artificially synthesizing one section of competitive internal amplification control and a corresponding probe,and establishing an internal standard fluorescent quantitation PCR system respectively, using ABI 7500 Software SDS 1.4 to analyze experiment results and taking amplification with a Ct value smaller than 36 as a detection positive result. According to the method, the method is provided with good specificity aimed at target species, false negative test results are avoided by monitoring PCR reaction in real time, a novel way is explored for identification of animal origin ingredients in food, and the method has the advantages of being accurate and stable, convenient to operate and the like.

The invention discloses a preparation method of liver cells with low expression or no expression of PERV. The preparation method includes the following steps: (1), respectively preparing a recombinant vector containing siRNA applied to RNA interference technology, a recombinant vector containing gRNA applied to CRIPRS/Cas9 technology and a vector containing Cas9 applied to CRIPRS/Cas9 technology; (2), jointly transfecting the three vectors to porcine somatic cells, culturing the transfected porcine somatic cells, and screening and identifying to obtain monoclonal genetically-modified porcine somatic cells; (3), utilizing somatic cell nuclear transfer technology to obtain genetically-modified pig containing nuclear genes of the genetically-modified porcine somatic cells; (4), extracting liver cells of the genetically-modified pig. According to the preparation method, knockout before transcription and inhibition after transcription of PERV genes are realized, and the technical scheme is effective and feasible.

The invention discloses a rice bacterial leaf blight resistance related gene OsABA2 which consists of nucleotide sequences of SEQ ID No.2 as shown in the specification. The invention further discloses a rice lesion mimic gene which is capable of remarkably improving the resistance of rice to bacterial leaf blight, and a lesion mimic mutant can be acquired through gene edition on the OsABA2 gene. Novel functions of the OsABA2 gene are discovered, and the lesion mimic mutant acquired after the OsABA2 gene is knocked off through gene edition can be applied to rice bacterial leaf blight resistance breeding; meanwhile, the lesion mimic trait is controlled by a single recessive nuclear gene, and hybrid identification and selection can be relatively simple to implement in transgenosis or crossbreeding; and in addition, the lesion mimic gene is derived from nonglutinous rice and has a great promotion function on bacterial leaf blight resistant breeding of the nonglutinous rice.

The method of breeding sweet pepper hybrid with nuclear two-purpose male sterility line of sweet pepper features the breeding process, which includes the cross-breeding, selfing and sib breeding process to transfer the male sterile nuclear gene (msms) into selfing sweet pepper line with excellent agricultural character to obtain nuclear two-purpose male sterility line of sweet pepper of sterile plant rate of 50 %; the agricultural character selectivity of sib breeding for 2 or 3 generations to selectively breed the two-purpose male sterility line of sweet pepper with stable agricultural character, stable sterile plant rate of about 50 % and sterile degree of 100 %; culturing the hybrid seed with the sterile plants of the nuclear two-purpose male sterility line as female parent and the selfing line as male parent, removing the breeding plants before pollination and collecting seeds of the female parent. The present invention uses the nuclear male sterility line as both sterile line and maintainer line to simplify the breeding process.

The invention provides a diagnostic kit for obesity gene mutation. Probe sequences are designed according to the principle of sequence reverse complementation in the direction from 5' to 3' specific to the coding sequence of obesity-related pathogenic genes, and the oligonucleotide in situ synthesis technology is adopted on a connecting joint of each probe sequence to conduct large-scale synthesisof oligonucleotide on a chip, the oligonucleotide on the chip is eluted with ammonia water to form an oligonucleotide mixture, and a biotin-labeled primer at 5' end is adopted to form a DNA probe library of biotin-labeled obesity-related pathogenic genes in a PCR method. A high-throughput screening method and application for obesity-related known sites are achieved, are faster, more economical and simpler than detection of single site-specific screening and whole-genome or whole-exon sequencing, have low equipment and environmental requirements, can be used in the polymorphic sites of obesity-related known nuclear genes, and is suitable for large-scale screening and preventive examination of obesity-related people.

The invention discloses a PCR (polymerase chain reaction) method and kit for identifying spina date seeds and counterfeits thereof on the basis of ITS sequence site. The invention provides primers for identifying spina date seeds, particularly a primer pair capable of amplifying amplification products of nucleotide disclosed as Sequence 1 in the sequence table and nucleotide disclosed as Sequence 2 in the sequence table. The experiment proves that screening is performed on the basis of the nuclear gene ITS sequence of the original plant to obtain the site specific primers for identifying the spina date seeds and counterfeits thereof, and the PCR reaction system and conditions are regulated to establish a site-specific PCR process. The site-specific primer PCR is simple to operate, and has the advantages of low cost, favorable repetitiveness, single identification strip, low sample consumption and the like. The specific primer PCR can greatly lower the false positive rate of PCR and ensure the identification accuracy.

The invention belongs to the technical field of molecular biological breeding, and relates to a PCR marker related to cabbage dominant nuclear gene male sterilityand application thereof. A method foridentifying or assisting in identifying the cabbage dominant nuclear gene male sterility is provided, it is detected that the genotype of to-be-detected cabbage C09 chromosome 29, 141, 514 positions is A/A or A/- or -/-, and the fertility of cabbage is determined according to the genotype of to-be-detected cabbage; the to-be-detected cabbage of the A/A genotype is normal; the to-be-detected cabbage of the A/- genotype is the heterozygous sterile; to-be-detected cabbage of the A/- genotype is homozygous sterile. By means of the PCR marker and a primer, the male sterility of the cabbage is assisted to be identified; the PCR marker has the advantages of being easy to operate, high in specificity and good in stability; the coincidence rate between identification results and field phenotypes reaches100%, early breeding of cabbage varieties can be achieved, and the PCR marker has great application prospects.

The invention discloses a method for obtaining male sterile lines of rice through fertility genes S44, and belongs to the technical field of biology. The method specifically includes the steps that amale sterile mutant controlled by single recessive nuclear genes is obtained through EMS mutagenesis rice, mutant character related genes are positioned through a Mutmap method, and the rice fertilityregulatory genes S44 are obtained. Mutation of the S44 genes can lead to a male fertility phenotype produced by rice, the fertility of plants can be regulated by regulating the expression of the S44genes, and according to the method for obtaining the male sterile lines of the plants and regulating the plant fertility, the great significance is achieved on researching anther developmental mechanisms of the plants and crossbreeding work of the rice.

The present invention provides production process of human lysozyme as AIDS treating medicine with plant as bioreactor. Human lysozyme gene is cloned from human placenta and constructed into prokaryotic expression vector for efficient expression. Plant expression vector is constructed in lettuce, and the human lysozyme gene is introduced into lettuce nuclear genome to realize the efficient expression of human lysozyme gene in higher plant and to obtain one kind of lettuce containing human lysozyme. Human lysozyme mass produced in the said method may be used in treating AIDS, tumor and viral diseases.

The invention belongs to a molecule detection method of Fusarium graminearum to medium resistance level bacterial strain of carbendazim. The method can be used for monitoring drug resistance of Fusarium graminearum to carbendazim and prevalence warning, wherein Fusarium graminearum can cause wheat scab. The detection method includes the following three main steps: (1) nuclear genome DNA of bacterial strain to be detected is respectively extracted: (2) internal and external primer pairs are utilized to carry out nested PCR, thus obtaining a target segment; (3) the target segment is respectively restricted by restriction enzymes HindIII and TaaI (Tsp4CI), and genotype of medium resistance level bacterial strain can be accurately identified by virtue of a restriction product electrophoresis spectrum. By adopting PIRA-PCR (primer-introduced restriction analysis PCR) technology, quantity of Fusarium graminearum with medium drug resistance in field and ratio thereof in group can be rapidly and accurately detected. Detection accuracy reaches more than 95%.

The invention discloses a method for identifying cucumber hybrid purity based on cucumber mitochondrial paternal inheritance characteristics by using a mitochondrial genome SSR marker technology, relates to cucumber mitochondrial genome SSR marker development as well as methods for respectively performing polymorphism screening on cucumber varieties of different ecotypes by using the developed cucumber mitochondrial marker and performing purity detection on cucumber F1 seeds by utilizing a polymorphism marker, belonging to the field of biotechnology breeding. According to the invention, the purity of cucumber hybrid first-generation seeds is detected by using a cucumber mitochondrial genome sequence development SSR marker, and false hybrid seeds and selfing seeds from other pollen sources on maternal cucumber plants are expected to be distinguished; relative to research in variety identification aspect based on a nuclear genome SSR marker technology, a novel detection method is provided by mainly utilizing the cucumber mitochondrial paternal inheritance characteristics, and a technical preparation is made for tracing paternal/maternal mitochondrial genomes in the hybridization process. The method also can be applied to detection of hybrid first-generation seeds of other paternal plant materials.

The invention discloses a molecular marker and application of a rapeseed recessive nuclear sterile gene segment. The steps are as follows: A. Positioning BnMs3 and Bnrf in the Brassica oleracea by controlling the location of the rapeseed recessive nuclear sterile gene BnMs3 and Bnrf A genetic interval of less than 0.2cM on the N19 and N7 linkage groups of rapeseed; B. The TapidorBAC library was screened by marker screening combined with Southern hybridization, and the target clones JBnB043L23 and JBnB089D05 containing BnMS3 and Bnrf were obtained; C. BnMs3 was obtained by the sequencing of the target BAC The nucleotide sequence of the candidate gene segment with Bnrf; D, the difference between the nucleotide sequence of the BnMs3 and the allelic recessive gene Bnms3 gene segment according to C step and the difference between BnRf and the allelic recessive gene Bnrf gene segment Differences between nucleotide sequences, development and design of primers, and development of molecular markers for the BnMs3 and BnRf sterile gene segments. The application of molecular markers in the molecular marker-assisted breeding of the recessive genetic male sterile line and the temporary protection line of Brassica napus makes the molecular marker-assisted transfer of the recessive genetic male sterile line and the temporary protection line of rapeseed faster, more accurate and more accurate. efficient.

The invention relates to oligonucleotide primers and a probe used for precise and quantitative detection of ovine-derived materials, and a kit comprising the primers and the probe. The invention further relates to a digital PCR detection method used for quantitative detection of ovine-derived materials. The method involves use of the specific oligonucleotide primers and the fluorescent mark probe specific to the mutton single-copy nuclear gene. The invention further relates to the application of the specific oligonucleotide primers and the fluorescent probe specific to the mutton single-copy nuclear gene in quantitative detection of ovine-derived materials. By means of the digital PCR detection method, the content of ovine-derived materials in a sample can be determined precisely and sensitively.

The current invention provides the promoter of the Zea mays nuclear gene encoding glutamine synthetase. Compositions comprising this sequence are described, as are plants transformed with such compositions. Further provided are methods for the expression of transgenes in plants comprising the use of these sequences. The methods of the invention include the direct creation of transgenic plants with the cytoplasmic glutamine synthetase promoter by genetic transformation, as well as by plant breeding methods. The sequences of the invention represent a valuable new tool for the creation of transgenic plants, preferably having one or more added beneficial characteristics.

The invention provides a creating method of chili S (RfRf) type cytoplasmic male sterility restoring lines. The method is characterized in that existing sterile line materials are used as the female parents, existing N (RfRf) materials are used as the male parents, the cytogenes of the female parents are taken and combined with the nuclear genes of the male parents to obtain target materials, the target-genotype intermediate materials are bred fast, and breeding reliability, accuracy and efficiency are increased. The creating method has the advantages that the method is simple in breeding procedure, objective traits and procedures of breeding can be controlled, breeding efficiency is increased greatly, the types and number of the bred restoring lines are increased greatly, and a new approach is provided for chili heterosis utilization and sterility line breeding and creating and the like.

Plants made resistant to insects by transforming their nuclear genome with two or more DNA sequences, each encoding a different non-competitively binding B. thuringiensis protoxin or insecticidal part thereof, preferably the toxin thereof.

The invention relates to oligonucleotide primers and a probe for accurate and quantitative detection of a dog-derived component. The invention also relates to a digital PCR detection method for quantitative detection of the dog-derived component. The method comprises a step of utilizing the specific oligonucleotide primers and probe directed at a dog-derived single-copy nuclear gene. The invention also relates to a digital PCR detection kit for quantitative detection of the dog-derived component. The kit comprises the specific oligonucleotide primers and probe for digital PCR detection of the dog-derived component. The invention further relates to application of the specific oligonucleotide primers and probe directed at the dog-derived component to quantitative detection of the dog-derived component. With the digital PCR detection method and kit of the invention, whether fresh meat and preliminarily processed meat products contain the dog-derived component or not can be specifically, sensitively and accurately detected.

The invention discloses a polymorphic primer of a cinnamomum camphora nuclear genome SSR molecular marker and application thereof, and belongs to the technical field of forestry molecular biology. Thenucleotide sequence of the primer is shown as SEQ ID NO.1 to SEQ ID NO.14. The polymorphic primer is applied to the application of variety identification of cinnamomum camphora, and genetic structureand resource genetic diversity analysis of ancient cinnamomum camphora groups. A method includes the steps of genome DNA extraction, PCR amplification, polyacrylamide gel electrophoresis and data analysis. 186 individuals of 18 ancient cinnamomum camphora groups are subjected to genetic structure and resource genetic diversity study; the possibly forming reasons are analyzed according to existinggenetic structure and resource genetic diversity patterns of the cinnamomum camphora; and theoretical basis is provided for the wild cinnamomum camphora resource protection and utilization. The repeatability of the SSR molecular marker is high, and the real level of the genetic diversity of the cinnamomum camphora can be comprehensively disclosed.

The invention relates to a quantitative detection method, composition and kit of donkey/horse-derived components in a donkey-hide gelatin colloidal liquid semifinished product or finished product, belonging to the technical field of molecular biology. The method comprises the following steps: designing donkey/horse universal primers and donkey/horse specific probes according to the donkey/horse house-keeping gene single-copy genes, extracting DNA (deoxyribonucleic acid) of a sample to be detected, carrying out TaqMan qPCR (quantitative polymerase chain reaction) on the donkey-hide gelatin colloidal liquid semifinished product or finished product to perform quantitative detection on donkey/horse-derived components, calculating the number of copies of the donkey/horse-derived nuclear genes in the unit mass of the donkey-hide gelatin colloidal liquid according to the standard curve and the Ct value of the sample to be detected, and calculating the proportion of the donkey/horse-derived components in the donkey-hide gelatin, thereby quantifying the donkey/horse-derived components in the donkey-hide gelatin sample. The method, composition and kit have the advantages of high specificity and high sensitivity, implement qualitative detection on the donkey/horse-derived components in the donkey-hide gelatin, and can also implement quantitative analysis, thereby providing technological popularization and application for quality control of donkey-hide gelatin enterprises.