235 results about "Molecular identification" patented technology

The invention relates to the technical field of plant genetic engineering, and particularly provides a rice endosperm tissue specific promoter pPROLM26 and application thereof. According to the invention, a tissue-specific promoter with a nucleotide sequence as shown in SEQ ID NO.1 is separated from a rice alcohol-soluble protein gene, the tissue-specific promoter is cut off for four times, and then the cut-off promoter fragment with an endosperm-specific promoter function as shown in SEQ ID NO.2-5 is obtained after molecular identification is carried out on the cut-off promoter fragment and an RUBY visual marker; the betaine content of the expression product can reach 2.5% of the dry weight of the seeds through determination, and the method has very high application value in the field of plant synthetic biology. The invention further discloses application of the promoter or the truncated promoter fragment in the aspects of improving crop seed quality, improving crop traits, cultivating new varieties of transgenic plants by using key elements of the promoter or expressing high value-added proteins by using a seed bioreactor and the like.

The invention provides a local livestock and poultry variety identification method based on an SNP marker, and relates to the field of variety identification, and the method comprises the following steps: S1, genotype data acquisition, S2, genotype data quality control, S3, variety identification standard library establishment, and S4, bloodline content evaluation. According to the method, on the basis of a standardized molecular identification analysis process of genomic SNP marker bloodline content evaluation, genotype information of a core SNP site is analyzed, so that whole analysis process processing from original genotyping data to variety identity judgment is realized; the method can provide standardized, convenient and efficient molecular detection technical support for protection and utilization of genetic resources of local livestock and poultry in China, germplasm innovation and industrial development.

The present invention relates to the field of plant genetic engineering technology, specifically providing a rice endosperm tissue-specific promoter, pPROLM26, and its applications. The present invention isolates a tissue-specific promoter with the nucleotide sequence shown in SEQ ID NO. 1 from a rice alcohol-soluble protein gene. This tissue-specific promoter is truncated four times and then molecularly identified with a RUBY visualization marker to obtain truncated promoter fragments shown in SEQ ID NOs. 2 to 5, each with endosperm-specific promoter function. The expression product has been measured to contain up to 2.5% betaine of the seed dry weight, demonstrating its high application value in the field of plant synthetic biology. The present invention further discloses applications of the promoter or truncated promoter fragments in improving crop seed quality, improving crop traits, cultivating new transgenic plant varieties using key elements of the promoter, or expressing high-value-added proteins using seed bioreactors.

The invention relates to a gene ZmILR1 for controlling the size of a corn kernel as well as a molecular identification method and application of the gene ZmILR1, and belongs to the technical field of plant genetic breeding and functional gene research. The ZmILR1 gene encodes an IAA-amino acid hydrolase, mutation of the IAA-amino acid hydrolase causes corn kernels to become smaller, and overexpression of the gene can create a corn material with increased kernel length, kernel width and hundred-kernel weight. A functional molecular marker A1A4 is developed according to the sequence difference of the mutant gene and a wild type gene at the DNA level, and can be used for rapid identification and screening of small-grain corn materials. The invention provides a theoretical basis and an application tool for analyzing a molecular mechanism of the corn grain size and developing molecular marker-assisted breeding and hybrid seed breeding, and provides important gene resources and technical support for high-yield and high-quality molecular breeding of corn.

The invention relates to the technical field of molecular detection, in particular to a specific primer combination for distinguishing enteromorpha from sibling species, application of the specific primer combination and a molecular identification method. The primer combination comprises an upstream primer SEQ1 and a downstream primer SEQ2, the primer sequence of the SEQ1 is as shown in SEQ ID NO.01, and the primer sequence of the SEQ2 is as shown in SEQ ID NO.02. The primer combination is designed based on the difference of genomes of two algae, has high specificity and sensitivity, can effectively avoid cross reaction, and ensures the accuracy of a detection result. Meanwhile, in combination with a real-time fluorescent quantitative PCR (qPCR) technology, the method can effectively quantify the abundance of enteromorpha in an environmental sample or a laboratory sample, and provides reliable technical support for early warning, traceability analysis and prevention and control strategy formulation of green tide outbreak.

The invention relates to a method for extracting genome DNA of plants rich in secondary metabolites and a buffer solution, and belongs to the technical field of molecular biology and botany. The method solves the technical problems of low DNA extraction efficiency, poor purity, easy degradation and the like when a traditional DNA extraction method is used for treating plant tissues rich in secondary metabolites such as alkaloid, polyphenol and the like. Comprising the following steps: adding a complexing agent such as polyethylene glycol or polyvinylpyrrolidone when grinding plant tissues in a liquid nitrogen environment; splitting by using a cell wall splitting buffer solution containing dithiothreitol and a nonionic surfactant; carrying out DNA release and extraction at 60-70 DEG C by adopting a CTAB (Cetyltrimethyl Ammonium Bromide) extraction buffer solution containing polyethylene glycol and papain; and then purifying and precipitating to obtain high-purity genome DNA (Deoxyribose Nucleic Acid). The method can effectively remove alkaloid, protein and other impurities, significantly improves the DNA yield and purity, and is suitable for genome sequencing, genetic resource protection, medicinal plant molecular identification and the like of plants with high secondary metabolites such as Stephania kwangsiensis and the like.

The invention belongs to the technical field of medicines, and discloses a near-infrared fluorescent probe taking an alpha, beta-unsaturated acryloyl ester unit as a response site of sulfydryl. The fluorescent probe has great advantages in the aspects of biological small molecule identification, protein marking and disease diagnosis, and is one of the most convenient and most popular tools for exploring a complex biological process. The fluorescent probe molecule capable of releasing red fluorescence after being activated by sulfydryl is obtained for the first time by connecting a fluorophore with an alpha, beta-unsaturated acryloyl ester unit. The fluorescent probe molecule disclosed by the invention can be used for specifically detecting sulfydryl in an organism, so that the aim of accurately monitoring the sulfydryl in a cell is fulfilled.

The invention relates to the technical field of plant genetic engineering, in particular to application of an algae NySAT2 gene in regulation and control of alfalfa crude protein and detergent fiber. According to the invention, a sequence after codon optimization of an NySAT2 gene coding region is obtained by using gene synthesis and codon optimization technologies, an algae NySAT2 gene overexpression vector is constructed, a target gene is introduced into an alfalfa genome by using an agrobacterium rhizogenes mediated genetic transformation method, and a transgenic plant is obtained by using a genetic transformation system. Molecular identification and phenotypic analysis show that on the premise of keeping normal growth vigor (plant height and biomass have no significant difference from those of a wild type), the crude protein content of a medicago sativa plant with the overexpressed NySAT2 gene is remarkably increased compared with that of the wild type medicago sativa plant, and meanwhile, the contents of acid detergent fibers and neutral detergent fibers are remarkably reduced, so that the content of the crude protein in the medicago sativa plant with the overexpressed NySAT2 gene is remarkably increased. The forage quality and the feeding value of the medicago sativa are effectively improved.

The application discloses a high-sweetness-amino-acid-content Flammulina velutipes variety and a MNP molecular identification method and application thereof. The Flammulina velutipes variety has excellent properties, the cap is not easy to open, the content of sweet amino acids is high, the diversified market demand is met, the factory annual bottle cultivation is suitable, and the Flammulina velutipes variety has a good application and popularization prospect. The MNP molecular fingerprint of the Flammulina velutipes variety 'Shangyan A111' is comprehensive multiple amplification and sequencing technology, sequence analysis of all marker sites of multiple samples can be performed at one time, compared with ISSR, RAPD, SSR and other molecular markers and mushrooming test, the MNP molecular fingerprint has the advantages of high throughput, multi-target, high sensitivity and high precision. The MNP molecular fingerprint of the Flammulina velutipes variety 'Shangyan A111' has specificity and specificity in identifying the Flammulina velutipes strain 'Shangyan A111'.

This invention relates to a rapid identification method Lactobacillus plantarum The method for identifying strains, belonging to the field of species and strain identification, is based on the differences in three DNA barcode sequences. Lactobacillus plantarum CGMCC NO: 26171 strain from Lactobacillusplantarum This method allows for rapid identification from other strains of the same species. Compared to traditional morphological identification methods, the standard gene sequence obtained is beneficial for the molecular identification of *Lactobacillus plantarum* strains, effectively shortening the identification time. Three pairs of DNA barcoding primers enable specific amplification of the test strain, allowing for rapid identification. Lactobacillus plantarum strains.

To provide a gene testing method and a gene testing kit for analyzing a variable region of a TCR or a BCR in single-cell analysis.SOLUTION: A method including a step of capturing mRNA eluted from a single cell using an RT probe immobilized on a solid-phase surface and including a common sequence, a cell identification barcode, a first molecular identification barcode, and an oligo(dT) sequence, a step of performing a reverse transcription reaction in a reverse transcription reaction solution containing a TSO using the mRNA as a template to generate cDNA having a sequence added to a 3' end, a step of degrading the mRNA, a step in which a primer for inserting a second molecular identification barcode binds to a constant region adjacent to a variable region of a TCR or a BCR and includes the second molecular identification barcode, a step of ligating cDNA extended from the primer with a ligase, a step of amplifying a fragment including the second molecular identification barcode and the variable region of the TCR or the BCR using a primer, a step of performing sequence analysis, and a step of associating sequences.SELECTED DRAWING: Figure 2

The invention relates to the technical field of crop molecular identification, in particular to a crop variety identification method based on a multi-allelic variation SNP (Single Nucleotide Polymorphism) marker. The variety identification method comprises the following steps: carrying out genotype detection on a to-be-detected crop based on a multi-allelic variation SNP marker set, comparing a detection result with a contrast variety genotype or fingerprint database, and judging the to-be-detected crop according to a comparison result, the multi-allelic variation SNP marker set is obtained by mining whole genome sequencing data of the representative material of the to-be-detected crop; the fingerprint database is obtained by performing hybridization sequencing on the crops to be detected based on the multi-allelic variation SNP marker set. The detection method provided by the invention can be used for authenticity identification, substantive derived variety identification, genetic relationship identification, germplasm resource genetic evaluation and breeding of crops, and provides a new thought and method for crop variety identification, variety right protection, germplasm resource evaluation and the like.

The present application relates to the technical field of molecular detection, and particularly relates to a specific primer combination for distinguishing Enteromorpha prolifera and related species, application and molecular identification method thereof. The primer combination comprises an upstream primer SEQ1 and a downstream primer SEQ2. The primer sequence of SEQ1 is shown as SEQ ID NO. 01, and the primer sequence of SEQ2 is shown as SEQ ID NO. 02. The primer combination of the present application is designed based on the genomic difference of two kinds of algae, has high specificity and sensitivity, can effectively avoid cross reaction, and ensures the accuracy of the detection result. Meanwhile, combined with the real-time fluorescent quantitative PCR (qPCR) technology, the method can effectively quantify the abundance of Enteromorpha prolifera in environmental samples or laboratory samples, and provides reliable technical support for early warning, tracing analysis and prevention and control strategy of green tide outbreak.

The invention belongs to the field of biological quarantine, and particularly relates to a method for separating and identifying pigeon-derived liver escherichia coli. Escherichia coli separation and identification are carried out on disease samples from Guangdong, Guangxi, Guizhou, Hainan and the like, and the epidemicity of the disease samples is determined; collecting livers of acute dead young meat pigeons, and separating strains; carrying out gram staining, physiological and biochemical identification and molecular identification on the separated strain; the growth and reproduction cycle and hemolysis characteristics of the strain are detected, 76 Escherichia coli infection occurs in 113 disease samples, and the infection rate is 67%. According to the method, laboratory diagnosis of infectious diseases of the meat pigeons is systematically completed through sample collection and separation, culture and identification of strains, it is determined that the young meat pigeons are infected by escherichia coli, and a foundation is laid for subsequent antibiotic-free green breeding of the meat pigeons resisting escherichia coli infection.

This invention provides a combination of SNP molecular marker primers for the molecular identification of soft-fiber areca nut germplasm resources and their applications, belonging to the field of SNP molecular marker technology. Based on the constructed core areca nut germplasm resources, this invention conducts genetic background association analysis on the phenotypic characteristics of fruit fibers and develops SNP molecular marker primers based on areca nut fiber-specific loci. The development of these SNP molecular marker primers includes four pairs of specific primers. This invention also provides a method for detecting soft-fiber areca nut germplasm resources using SNP molecular marker primers. This method can rapidly, accurately, cost-effectively, and with high throughput detect the genotype of soft-fiber areca nut germplasm resources. The four pairs of specific primers of this invention can be used to identify soft-fiber areca nut germplasm resources, which has important application significance for the scientific protection of areca nut germplasm resource diversity, research on the genetic conservation and evolution of areca nut resources, and areca nut genetic breeding.

The application discloses a kind of molecular identification methods of sex-linked luhua feather homozygote of luhua chicken, belong to molecular marker assisted breeding technical field.The application can quickly and accurately detect homozygous sex-linked luhua chicken individual by designing KASP detection primer for detecting three sex-linked luhua feather color strong linkage sites, and the genotype of three sites can quickly and accurately detect homozygous sex-linked luhua chicken individual, when SNP2 (C->A) at 389bp of the first intron of CDKN2A gene, SNP3 and SNP4 at 172bp (T->A) and 174bp (T->C) of the first exon are AA, AA and CC respectively, then the chicken to be measured is homozygous sex-linked luhua chicken, otherwise the chicken to be measured is heterozygous sex-linked luhua chicken or non-sex-linked luhua chicken.The application provides an effective technical means for purifying sex-linked luhua feather color gene of luhua chicken, and helps to speed up the breeding speed of sex-linked luhua chicken strain homozygote and its matching system.

This invention belongs to the field of environmental microbiology technology, specifically disclosing a *Aspergillus niger* strain with lead adsorption capacity and its applications. This invention involved the isolation of microorganisms from 33 samples of water and sediment from the Henan section of the Yellow River basin. A fungal strain with high lead tolerance and adsorption was screened out, and after morphological and ITS molecular identification, it was named *Aspergillus niger* AN-2 and deposited at the Guangdong Provincial Microbial Culture Collection Center (GDMCC) with accession number GDMCC NO: 63697. Compared with other isolated strains, *Aspergillus niger* AN-2 exhibits high lead tolerance and adsorption capacity for Pb. 2+ It has the strongest tolerance and is most tolerant to Pb. 2+ The MIC is as high as 3500-3800 mg / L. When this strain is processed into biomass, it exhibits excellent adsorption performance for lead, making it suitable for preparing bioremediation agents for heavy metal pollution. It has broad application prospects and high economic value.

The invention discloses a KASP molecular marker primer group for identifying saline-alkaline tolerance of rape and application of the KASP molecular marker primer group, and belongs to the technical field of molecular marker-assisted breeding. Comprising a first primer (SEQ ID NO.2), a second primer (SEQ ID NO.3) and a universal primer (SEQ ID NO.4), the primer group is designed based on a key SNP site (A / T) of a BnaA02. CDKE1 gene, and rapid and accurate typing of rape genotypes can be realized through a KASP technology. The invention also provides a kit containing the primer group, an application of the kit in saline-alkaline resistance identification of rape, and a corresponding identification method. The method has the advantages of simplicity and convenience in operation, stable result, high flux, low cost and the like, and is suitable for large-scale auxiliary breeding and molecular identification of saline-alkaline tolerant rape varieties.

This invention relates to nucleic acid aptamers for lipoprotein-associated phospholipase A2 (Lp-PLA2), wherein the nucleic acid aptamer is at least one of B76-2, B76-4, and B76-5, with sequences shown in SEQ ID NO.1, SEQ ID NO.2, and SEQ ID NO.3, respectively. The Lp-PLA2 aptamers provided by this invention fill a gap in the field of Lp-PLA2 aptamers and exhibit high affinity and specificity. Three aptamers with good affinity and specificity for Lp-PLA2 have been screened and verified. These aptamers can be formulated into molecular probes or used as detection reagents in methods or kits for detecting Lp-PLA2, improving accuracy. They can also be used to develop novel Lp-PLA2 biosensing methods for molecular identification, showing great application potential in the early warning of cardiovascular disease events.

The application provides a SNP molecular marker related to soybean seed oil content, protein content and hundred-seed weight and application. The SNP molecular marker contains a nucleotide sequence with C / T polymorphism at the 201th position of the sequence shown in SEQ ID NO:1. The molecular marker of the application has very important guiding significance in soybean molecular identification and assisted breeding.

The invention discloses a multi-label smell description prediction method, and relates to the field of compound smell prediction, and the method comprises the steps: obtaining compound identification information, molecular structure descriptors and smell label data, and constructing a multi-label smell data set; generating a molecular structure feature vector through a molecular fingerprint coding technology, and extracting a multi-dimensional descriptor reflecting the physicochemical properties of molecules; compressing the molecular fingerprint features to a low-dimensional space through a dimension reduction algorithm; performing unbalanced data processing on the training set, fusing the dimension-reduced molecular fingerprints with the molecular descriptors to form a joint feature matrix, and configuring a class weight balance mechanism and overfitting suppression parameters by adopting a multi-label classification architecture; independently optimizing a probability threshold for each odor label based on the verification set; and outputting a multi-odor label combination prediction result according to the target molecule identification information. According to the scheme, the multi-odor characteristics of the compound can be accurately depicted, and the combined recognition accuracy of the compound odor is remarkably improved.

The application discloses the technical field of molecular identification of plant species, and discloses molecular markers for identifying five species of Chimonanthus and application of the molecular markers. The application develops specific molecular markers (cox2i691 and nad4i1399) based on variable regions between mitochondrial genomes of Chimonanthus praecox and Chimonanthus nitens, and effectively distinguishes the five species of Chimonanthus, i.e. Chimonanthus praecox, Chimonanthus nitens, Chimonanthus salicifolius, Chimonanthus zhejiangensis and Chimonanthus oblongus. The application is beneficial to conveniently and accurately identifying the species of Chimonanthus, and provides important information for analyzing evolution of the Chimonanthus and analyzing a forming mechanism of biochemical properties in the future, and is also beneficial to classification of the species of Chimonanthus, evolution research of mitochondrial genomes, and product discrimination of the Chimonanthus species.

The invention relates to the technical field of molecular identification of muskmelon hybrid seed purity, discloses an SSR primer and a method for rapidly identifying muskmelon hybrid seed purity, and aims to solve the problems that traditional seed purity detection depends on field test and manual observation, the detection period is long, the cost is high and the like. The invention provides two SSR primers with high polymorphism and good specificity for the melon hybrid 'Suqian No.4' and the specific markers thereof, and the SSR primers are applied to purity identification of 'Suqian No.4' seeds, so that good technical support is provided for seed production and seed sales, and the result is reliable. The method has the advantages of rapidness, accuracy, low cost, good repeatability, strong operability and the like.

The invention relates to the technical field of gene editing, and particularly provides a method for creating fluazifop-p-butyl herbicide resistant rice through gene editing of an OsACCase gene promoter region. The method comprises the following steps: constructing a crRNA plasmid library covering an OsACCase gene promoter region on the basis of a CRISPR / LbCas12a system, performing agrobacterium-mediated transformation on rice callus, and performing resistance screening, fluazifop-p-butyl herbicide screening and molecular identification to obtain a fluazifop-p-butyl resistant rice strain. The strain is subjected to-4bp / -5bp double allelic deletion mutation in an upstream 1865-site region of an initiation codon ATG of the OsACCase gene, and a homozygous mutant strain and a double allelic deletion mutant strain can normally grow after being sprayed with 6 times of field recommended dose of fluazifop-p-butyl. The herbicide resistance is realized by editing the gene promoter region, the growth defect of coding region mutation is avoided, the breeding technology is enriched, and weed prevention and control are assisted.

The present invention discloses a method for molecular identification of crop germplasm resources and its application, which solves the technical problem that the existing genetic markers are used to classify and identify rice germplasm resources. Due to the relatively few polymorphic markers, the identification results are often not ideal, and there is a lack of a method for quickly and accurately identifying germplasm resources at the molecular level. The present invention includes the following steps: Step 1: Using the differences in agronomic traits, screening gradient classification markers; Step 2: Using the screened gradient markers to detect germplasm resources; Step 3: According to the detection results of the gradient markers on crop germplasm resources, evaluating the genetic relationship of the resources. The present invention has the advantages of making up for the deficiencies of traditional phenotypic identification techniques and increasing the accuracy and reliability of resource identification.

The invention provides an EST-SSR (expressed sequence tag-simple sequence repeat) primer group for astragalus sinicus and application, and belongs to the technical field of molecular identification. The EST-SSR primer provided by the invention realizes high specificity, polymorphism and amplification stability through precise design based on astragalus sinicus transcriptome data in combination with a two-step screening process, can effectively distinguish genetic differences of different astragalus sinicus varieties, and can be used for solving the problem of intra-variety heterozygosis caused by cross pollination. The technical system provided by the invention has economical efficiency and practicability, an efficient and reliable tool is provided for germplasm resource identification, genetic diversity analysis and molecular breeding of astragalus sinicus through low-cost and high-throughput fluorescent SSR markers, and the technical blank of molecular marker development of green manure crops is filled up; the method has important application value for genetic improvement and resource management of astragalus sinicus.

The application discloses a molecular marker for identifying salvia miltiorrhiza cultivars and application thereof, and belongs to the technical field of plant molecular identification. The molecular marker comprises SM-M1, SM-M2, SM-M3, SM-M4, SM-M5 and SM-M6. The molecular marker can be used for effectively identifying two subgroups of salvia miltiorrhiza cultivars, and is beneficial to the research on genetic variation in salvia miltiorrhiza and evolution of cultivars.

The invention discloses a rumen fibroblast line as well as a construction method, a culture method and application thereof, the rumen fibroblast line G8 is preserved in the China Center for Type Culture Collection, the preservation number is CCTCC NO: C2025332, the rumen fibroblast line G8 has a typical fibroblast-like form and can be stably passaged, and the rumen fibroblast line G8 can be used as a rumen fibroblast cell line. The cell culture medium also has the characteristic of highly expressing fibroblast marker genes and the function of tolerating and utilizing rumen core metabolite butyric acid, and the proliferation rate and metabolic activity of the cell culture medium under the condition that 15mmol / L butyric acid is used as a main energy source are obviously superior to those of glucose culture; according to the construction method of the rumen fibroblast line, the steps of function screening and molecular identification are introduced, so that the identity authenticity of the constructed cell line can be ensured; the culture method of the rumen fibroblast line is simple and effective, culture medium materials are easy to obtain, and culture conditions are easy to realize; the rumen fibroblast line can be used for rumen in-vitro research, and is beneficial to clarification of a mechanism for regulating rumen interstitial metabolism and tissue repair by butyric acid.

The invention relates to a mycelium extracted from ganoderma lucidum and a preparation method of a culture medium of the mycelium, belongs to the technical field of mycelium extraction, and aims to solve the problems that in the prior art, ganoderma lucidum mycelia grow slowly and are prone to pollution, the content of effective components is unstable, and molecular identification is difficult. The invention provides the solid culture medium prepared from the saussurea involucrata purple yellow extract, the extract is formed by combining effective components of saussurea involucrata and sodium alginate, the growth rate and metabolism efficiency of ganoderma lucidum hyphae can be improved, active substances such as polysaccharides and triterpenes are effectively enriched, and the yield of the ganoderma lucidum hyphae is increased. Meanwhile, the stress resistance and the survival rate of the mycelium under the stress conditions of oxidative stress, pathogen invasion and the like are enhanced, the pollution risk in the culture process is reduced, the biosynthesis efficiency and the extracellular secretion level of key secondary metabolites are improved, the immunocompetence and the bioavailability of the product are further improved, and the production cost is reduced. And a stable and reliable technical support is provided for large-scale and high-quality production of ganoderma lucidum.

The present application belongs to the technical field of molecular identification of new shrimp species, and particularly relates to a composition for identifying Neocaridina denticulata and Neocaridina zhoui and use thereof. The present application discloses a composition for identifying Neocaridina denticulata and Neocaridina zhoui, which comprises: a first specific primer pair with a nucleotide sequence as shown in SEQ ID NO: 3 and SEQ ID NO: 4; and a second specific primer pair with a nucleotide sequence as shown in SEQ ID NO: 5 and SEQ ID NO: 6. Based on the composition and the corresponding duplex PCR method provided by the present application, a 743 bp band specific to Neocaridina denticulata or a 601 bp band specific to Neocaridina zhoui can be amplified simultaneously in one reaction system, so that the two morphologically similar new shrimp species can be identified quickly, accurately and efficiently.