90 results about "Histone" patented technology

The invention relates to the technical field of biology, in particular to a molecular marker for accurately positioning centromere of chromosome of Chinese trumpet creeper, and the sequence of the molecular marker is as shown in SEQ ID NO.1 and SEQ ID NO.2. The method comprises the following steps: specifically enriching a DNA (Deoxyribose Nucleic Acid) sequence combined with a centromere specific histone CENH3 through a chromatin co-immunoprecipitation technology (ChIP), and sequencing the DNA sequence; sequencing data are analyzed to obtain a centromere sequence of the vicia villosa, and two specific repetitive sequences of the centromere of the vicia villosa are obtained after identification and optimization. Primers are designed according to the sequences, PCR amplification is carried out, amplification product sequences are marked, fluorescence in situ hybridization verification is carried out by analyzing a cytogenetics technology, it is proved that the primers are specifically distributed in a centromere area of vicia villosa, and the centromere specificity of the primers is proved. The marker can be used for genetic identification and accurate positioning of the centromere in genome research, and can also be used for cytological identification of the centromere and karyotype research such as chromosome counting and morphological analysis through centromere signal observation.

The invention discloses a method for screening and identifying a cucumber aphid-resistant gene as well as a screened gene and application thereof, and provides a brand-new method for screening and identifying the cucumber aphid-resistant gene, which is used for screening the cucumber aphid-resistant gene based on four omics technologies. The four types of omics comprise transcriptome, proteome, methylation group and metabolome before and after inoculation of aphids on cucumbers, and the cucumber aphid resistance regulation gene is screened through combined analysis, so that a new gene resource is provided for improving the aphid resistance of the cucumbers, and a new method and a new idea are provided for designing molecular genetic breeding. The CsPGD2 gene obtained through screening is an aphid resistance gene, the dynamic change of a metabolic substance 5-ribulose phosphate in a metabolome is consistent with that of a transcriptome gene CsPGD2, the aphid resistance function of the gene CsPGD2 is verified through VIGS, and the result proves that the gene CsPGD2 is an important regulation factor for the aphid resistance of cucumbers. The invention provides a new gene resource for improving the aphid resistance of cucumbers, and has a good application prospect.

The present invention relates to the technical field of biomedicine. Specifically provided are a respiratory syncytial virus (RSV) Pre-F recombinant protein nanoparticle vaccine and a preparation method therefor. By performing amino acid mutation on an RSV Pre-F protein, and performing fusion expression on the mutated Pre-F protein and ferritin particles in eukaryotic cells, PreF-Ferritin fusion protein nanoparticles, in which eight Pre-F protein trimers are densely displayed on the surface of each ferritin particle, are obtained. By stabilizing and exposing antigen epitopes required to be displayed and disrupting or masking unrequired antigen epitopes, the PreF-Ferritin fusion protein nanoparticles effectively improve the immunogenicity and the production stability of the antigen. Experiments show that: when the PreF-Ferritin fusion protein is injected into mice, high-titer protective sera can be acquired, and the mice sera can yield relatively high neutralizing titers against RSV.

Metastatic uveal melanomas are highly resistant to all existing treatments. To identify actionable vulnerabilities, the inventors conducted a CRISPR-Cas9 knockout screen using a library composed of chromatin remodelers. They revealed that the histone H3 methyltransferase SETDB1 plays a critical role in metastatic uveal melanoma cell proliferation and survival. Functionally, SETDB1 knockdown triggers decreased expression of genes related to replication and cell cycle and promotes growth arrest associated with increased markers for DNA damage and senescence entry. Using pre-clinical model, they further demonstrated that anti-SETDB1 therapy tumor growth in vivo. The inventors identify SETDB1 as a new relevant therapeutic target for the treatment of metastatic uveal melanomas. The present invention relates to a method for treating uveal melanoma in a subject in need thereof comprising a step of administering said subject with a therapeutically effective amount of SETDB1 inhibitor.

The invention discloses an efficient extraction method of blue fox sperm DNA, and belongs to the technical field of molecular biology. Aiming at the structural characteristics that chromatin of the blue fox sperms is highly condensed, histone is replaced by protamine, and a stable nucleoprotein complex is formed through an intermolecular disulfide bond, the invention provides an efficient DNA extraction method suitable for the blue fox sperms. According to the method, based on the synergistic effect of three chemical reagents, namely SDS, PK and DTT, full lysis of the sperm cells of the blue foxes is achieved, and then efficient enrichment of sperm DNA of the blue foxes can be achieved by combining a conventional DNA extraction method. According to the efficient extraction method of the blue fox sperm DNA provided by the invention, the yield and integrity of the blue fox sperm DNA are remarkably improved, and the limitation that sperm chromatin is difficult to effectively lyse by a conventional method is overcome; the method provides reliable technical support for research on genetic diversity evaluation, population management, molecular breeding and the like of the blue foxes, and has the potential of popularization and application in other high-condensation sperm species.

The invention relates to a marker for predicting the prognosis of ovarian cancer and application thereof, and the marker is used for predicting the prognosis condition of ovarian cancer by detecting any one or more of BRD4, H4K12la and Ube2v1. Compared with the prior art, it is found for the first time that BRD4 can be used as a histone milk acylation transferase to enzymatically modify a histone lysine site H4K12, so that the histone lysine site H4K12 is subjected to milk acylation modification, then gene transcription of Ube2v1 is increased, and proliferation and growth of ovarian cancer cells are promoted. The invention not only discovers that BRD4 can be used as histone milk acylation transferase to play a new function of milk acylation modification, but also clarifies the effect of the BRD4-H4K12la-Ube2v1 pathway in the growth process of the ovarian cancer, and provides a new method for targeted therapy of the ovarian cancer.

The invention relates to the technical field of biological information, and particularly discloses a multi-omics genome combined genetic assessment method and device. According to the method, multi-source data such as a genome, a transcriptome, a proteome, a metabolome, a microbiome and a high-throughput phenotype are integrated by constructing a multi-omics fusion similar matrix, an adaptive dichotomy is adopted to optimize omics weights, a grid search and cross validation system is introduced, and global optimal estimation of parameters is achieved. Verification of the method on livestock and poultry breeding and plant breeding data shows that the prediction accuracy of complex characters can be remarkably improved, and particularly, the prediction precision in low heritability characters is remarkably improved. The invention further provides a corresponding device which comprises a data acquisition module, a matrix construction module, a parameter optimization module and a breeding value calculation module, comprehensive utilization of multi-omics information can be efficiently achieved, and the accuracy and calculation efficiency of breeding value estimation are improved.

The invention discloses a gene level DNA methylation, transcriptome and proteome conjoint analysis method based on a snkemake framework, which comprises the following steps: preprocessing original data of DNA methylation and / or transcriptome and / or proteome, analyzing gene methylation, analyzing transcriptome and / or proteome, and analyzing the transcriptome and proteome. Based on gene methylation analysis data, the preprocessed transcriptome data and the preprocessed proteome data, correlation analysis and / or difference intersection analysis and / or enrichment pathway conjoint analysis are / is carried out, finally analysis results are sorted, and a visual report is generated. The method has comprehensive results, and relates to quantification, difference analysis and pathway enrichment analysis of gene methylation, and correlation analysis, difference intersection analysis and enrichment pathway joint analysis of integrated DNA methylation and / or transcriptome and / or proteome data; automatic arrangement, verification, visualization and report generation of analysis results are realized; all operation steps can be traced, and corresponding analysis log records are generated. The invention further discloses a related system and application.

The invention provides an expression vector, an expression system and a preparation method of a recombinant larosidase protein, and belongs to the technical field of recombinant protein preparation, the recombinant expression vector of the larosidase protein provided by the invention comprises an initial vector and a codon-optimized human-derived larosidase coding gene; the nucleotide sequence of the human-derived larosidase coding gene is as shown in SEQ ID NO. 1; according to the recombinant expression system, Chinese hamster ovary cells are taken as host cells, and the recombinant expression vector is transfected. According to the invention, a gene sequence more suitable for expression of Chinese hamster ovary cells is obtained through codon optimization, and then transposon-mediated stable integration is carried out; high-density suspension culture and downstream purification processes are utilized to realize efficient and stable expression of the recombinant larosidase in CHO cells, and the recombinant larosidase with high yield, high activity and high purity can be obtained.

The present disclosure includes, among other things, methods, kits, and systems for determining a status of lung cancer. In various embodiments, the present disclosure relates to the use of one or more histone modifications, chromatin accessibility, binding of one or more transcription factors, and / or DNA methylation as features of a status of lung cancer. In some embodiments, differential modifications and / or differential accessibility are detected and quantified at one or more genomic loci in a biological sample, e.g., cell-free DNA (cfDNA), from a liquid biopsy sample obtained or derived from a subject having lung cancer. In various embodiments, the determined status can be used, e.g., to select a treatment for lung cancer and / or to treat lung cancer.

The invention belongs to the technical field of gene engineering, and particularly relates to a CHO cell transient transfection method and application thereof.The CHO cell transient transfection method comprises the following steps that subcultured CHO cells are resuspended through a transfection culture medium, the cell density in CHO cell suspension is 0.8 * 10 < 7 >-1.5 * 10 < 7 > cells / mL, a PEI transfection reagent containing 40 kDa linear PEI is used, plasmid DNA and the PEI transfection reagent with the mass ratio being 1: (1-6) are added into the CHO cell suspension for transfection, and transfection is conducted after the PEI transfection reagent is added into the CHO cell suspension; the addition amount of plasmid DNA in each milliliter of CHO cell suspension is 3.0-7.0 [mu] g, transfecting for 20-24 h, then adding a cell suspension culture medium, transfecting for 48-72 h, then repeatedly transfecting, and culturing to obtain the recombinant CHO cell. According to the method, the transfection efficiency can be improved, more cells for expressing the recombinant protein can be obtained, and the transient expression of the target protein can be remarkably improved.

The invention provides a histone demethylase gene HOVUSG0811000 related to powdery mildew resistance and an application of the histone demethylase gene HOVUSG0811000 related to powdery mildew resistance. The nucleotide sequence of the HOVUSG0811000 is as shown in SEQ ID NO. 1 (sequence identifier number 1). It is found for the first time that HOVUSG0811000 improves the resistance of highland barley to powdery mildew by regulating phenol amine metabolism, and the HOVUSG0811000 has a good application prospect in construction of powdery mildew resistant plant varieties.

The present disclosure includes, among other things, methods, kits, and systems for determining the status of MS in a subject. In various embodiments, the present disclosure relates to the use of one or more histone modifications, chromatin accessibility, binding of one or more transcription factors, and / or DNA methylation that are characteristic of the status of MS. In some embodiments, differential modifications and / or differential accessibility are detected and quantified, at one or more genomic loci of a biological sample, e.g., in cell-free DNA (cfDNA) from a liquid biopsy sample obtained or derived from a subject with MS. In various embodiments a determined status is useful, e.g., in selecting treatment for and / or treating MS.

The invention discloses a novel nuclear magnetic resonance contrast agent Gd-DOTA-PP for targeting citrullinated histone to achieve neutrophil extracellular trap net imaging and a preparation method and application thereof.The method comprises the steps that citrullinated histone targeting polypeptide PP and ligand molecules DOTA-NHS are combined, and a compound DOTA-PP is obtained; and then gadolinium (Gd) ions are introduced into the DOTA-PP through a coordination reaction, and the novel contrast agent Gd-DOTA-PP is prepared. The contrast agent can be selectively combined with an important marker, namely citrullinated histone, of neutrophil extracellular trapping nets (NETs), and T1 weighted imaging signals of a target area are remarkably enhanced. The preparation method is simple and convenient to operate, the preparation process is highly controllable, and the obtained contrast agent has excellent biocompatibility and targeting characteristic. The accurate targeting ability of the probe enables the influence signal of the probe in NETs enrichment areas such as tumor early metastasis, tumor infiltration lymph nodes and the like to be obviously enhanced, a powerful tool is provided for tumor diagnosis, especially accurate diagnosis of the early metastasis, and the probe has important clinical application value.

The invention discloses application of an epiregulatory factor gene in regulation of plant heat resistance. The epiregulatory factor gene (TaMET) is identified from a wheat genome for the first time. The gene is subjected to transient overexpression in wheat Fielder, and it is found that wheat can be more sensitive to heat expression. The homologous gene (OsMET) of the gene in rice is subjected to conservative structural domain analysis to find that the gene is an SAM dependent type methylated transferase gene, and the modification level of histone H3K27me3 can be influenced. Research finds that overexpression of the gene makes the plant more sensitive to heat, knockout of the gene can significantly improve the heat resistance of rice Nipponbare, that is, the gene belongs to a negative regulation factor of rice heat resistance, for example, deep analysis of a mechanism of the gene or a homologous gene thereof is beneficial to excavation of heat-resistant genes in rice and wheat.

The application discloses a transient transfection method for improving recombinant protein expression of CHO cells, and relates to the technical fields of eukaryotic cell gene transfection and animal cell culture. A cell suspension is obtained by mixing a CHO cell and a serum-free CHO cell basic culture medium; DNA and a transfection reagent are added into the cell suspension to mix a transfection culture, 5-azadeoxycytidine acid solution is added into the transfection culture at 0h-24h to form a transfection system and continue the transfection culture; the transfection system is added with a feeding culture medium at 1d-4d after the transfection culture to carry out suspension culture; the suspension culture is centrifuged at 7d to separate and harvest cell culture supernatant to complete recombinant protein expression. According to the application, 5-azadeoxycytidine acid is added into the transfection culture at a specific concentration, so that the expression of recombinant protein of the CHO cell is significantly improved during the transfection expression.

The present invention provides a hybridoma cell line secreting monoclonal antibody against grass carp IL-4 / 13B, the monoclonal antibody and its application. The preservation number of the hybridoma cell line is CCTCC NO: C2022213. The monoclonal antibody against grass carp IL-4 / 13B is secreted by the hybridoma cell line, and the monoclonal antibody specifically recognizes the prokaryotic expression product, eukaryotic expression product and extracellular expression functional region protein of grass carp IL-4 / 13B. The preparation method of the grass carp IL-4 / 13B recombinant protein in the present invention is simple and highly expressed in animal cells. The expressed grass carp IL-4 / 13B protein can be used to find protein molecules that can activate or inhibit grass carp IL-4 / 13B, and can also be used to prepare monoclonal antibodies against grass carp IL-4 / 13B. The antibody can be used to prepare reagents and drugs for diagnosing, treating or preventing diseases related to the expression of grass carp IL-4 / 13B protein.

The invention discloses sodium polystyrenesulfonate modified chitosan as well as a preparation method and application thereof, and relates to the technical field of biomedical materials. The modified chitosan provided by the invention can generate far better adsorption capacity with the histone by virtue of weak amino positive electricity, so that the adsorption of the chitosan on the damage-related molecular mode of the histone is realized, and the long-standing technical bottlenecks of poor selectivity, low efficiency and the like in the field of histone separation and adsorption are solved; and the preparation method is simple and high in biological safety, and has relatively strong technical frontier and market application potential.

A small activating RNA (saRNA) capable of activating a CCAAT enhancer binding protein alpha (CEBPA) gene, and a delivery system and use thereof are provided, where sense and antisense strands of the saRNA have nucleotide sequences set forth in SEQ ID NO: 1 to SEQ ID NO: 2, respectively. A biomimetic nano-delivery system for targeted delivery of the saRNA is a biomimetic nanoparticle formed by composite nanoparticle coated by a biomembrane of an inflammatory effector cell, where the composite nanoparticle is formed by histone loading with the saRNA capable of activating the CEBPA gene. The biomimetic nanoparticle can inherit antigens and related membrane functions of the inflammatory effector cell and specifically accumulate at the sites of inflammatory lesions, which enables the biomimetic nanoparticle to not only have a longer circulation time in vivo, but also show inflammatory tropism.

The application discloses an ELISA kit for detecting anti-chromatin antibodies. The kit contains purified chromatin monomers with stripped H1 histone components in a natural structure. The application also discloses a detection method for the ELISA kit. The chromatin structure in the eukaryotic cell nucleus is complex, and is a natural complex composed of 40% DNA, 40% histone, 20% non-histone, a small amount of RNA and other macromolecules. The purification technology is high in requirement, and the price of the natural purified chromatin monomers is high. The application purifies the chromatin monomers with stripped H1 histone components according to the structure of the natural chromatin, retains the effective antigen components, and greatly reduces the cost. The kit can be used for detecting the anti-chromatin antibodies, fills the blank of the domestic anti-chromatin antibody detection method, and has the characteristics of large detection flux, accurate result, high sensitivity and simple operation.

The present disclosure includes, among other things, methods, kits, and systems for determining the status of lung cancer. In various embodiments, the present disclosure relates to the use of one or more histone modifications, chromatin accessibility, binding of one or more transcription factors, and / or DNA methylation that that are characteristic of the status of lung cancer. In some embodiments, differential modifications and / or differential accessibility are detected and quantified at one or more genomic loci of a biological sample, e.g., in cell-free DNA (cfDNA) from a liquid biopsy sample obtained or derived from a subject with lung cancer. In various embodiments a determined status is useful, e.g., in selecting treatment for and / or treating a lung cancer.

Disclosed are liquid and solid assay compositions and portable sample reader devices for use in a sample-to-answer diagnostic system for the detection of one or more analytes, preferably the detection of circulating histones in whole blood. Further provided are methods of making and using the assay compositions and portable sample reader, including the collection of a raw sample, testing the sample using the assay compositions, and analyzing the sample via the portable sample reader. More particularly, assay compositions comprising a sacrificial partition, target molecule, detectable label, and sacrificial partition are used in combination with a sample reader comprising an optical system and a housing unit as part of a sample-to-answer diagnostic system for quantifying circulating histones in whole blood as a mechanism of predicting the risk of multiple organ failure.

The present discloses provides the use of tributyrin as an additive for embryonic development culture medium in vitro, in the present disclosure, the tributyrin is applied as an additive to the development culture medium of mouse embryos in vitro for the first time, the tributyrin can significantly increase the rate of blastocyst, and reduce the ROS content in the embryos, improve the mitochondrial membrane potential in the embryos, increase the ATP level and the expression of antioxidant genes in the embryos, improve the DNA methylation and histone modification level in the embryos, promote the embryos development in vitro; In addition, tributyrin, as natural antioxidant and apparent drug, which is safe, non-toxic and side effects; the tributyrin provides strong support for the efficient embryos development of human assisted reproductive technology, mammalian fertilization embryos, parthenogenetic embryos and somatic cell cloned embryos and other embryo engineering technologies in vitro.

The invention discloses a method for capturing bacteria through a bionic micro-grid and application of the method, and belongs to the technical field of biological medicine. According to the method, a neutrophil extracellular trap (NETs) mechanism is simulated, salmon sperm DNA and calf thymus histone are subjected to ultrasonic complexing, and a bionic micro-net material (mu web) with a porous net structure is prepared. According to the micro-grid, the contact area of bacteria is increased through a net structure, the bacteria are efficiently captured through charge interaction, and the micro-grid has the advantages of being high in enrichment efficiency and low in biotoxicity. By adjusting the ratio of histone to DNA, the bacterium capture performance of the micro-grid can be optimized. The material can be applied to enrichment of corpse microorganisms in water in forensic practice and separation of low-concentration pathogenic bacteria in food safety detection, and a new way is provided for enrichment of bacteria in complex samples.

This invention relates to the fields of gene editing and animal genetic breeding technology, and discloses a precise editing method for key genes regulating lipid metabolism in cattle and sheep. The method includes: constructing a comprehensive lipid metabolism network map integrating transcriptomic, proteomic, and metabolomic data; using a graph neural network to identify key node genes with high flow weights and their negative feedback pathways; simulating cascade reactions after gene perturbation to assess compensatory effects; screening for multi-gene combinations that do not induce significant compensation and can synergistically regulate fat deposition and muscle growth; designing expression vectors containing multiple guide RNA sequences to achieve simultaneous and precise editing; and obtaining edited individuals and validating the phenotype through embryonic or somatic cell nuclear transfer. This invention, through system-level multi-target synergistic editing, avoids metabolic imbalances, improves editing effectiveness, and promotes muscle development while reducing fat deposition, providing a safe and efficient technical path for precision breeding of cattle and sheep.

This invention discloses self-assembled ferritin nanoparticles and their application in the preparation of duck hepatitis A vaccine. The invention connects the VP1 portion of the duck hepatitis A virus P1 protein to the N-terminus of a self-assembled ferritin nanoparticle subunit via a linker sequence to obtain a fusion protein, the amino acid sequence of which is shown in SEQ ID NO.1. To improve the expression level or efficiency of the fusion protein in silkworms, the nucleotide sequence of the encoding gene of the fusion protein is optimized and modified according to the codon preference of silkworms, resulting in the optimized gene sequence shown in SEQ ID NO.2. The invention also involves single-site or multi-site mutations of the fusion protein, screening for mutants with significantly improved titers and significantly increased soluble expression levels. This invention utilizes a silkworm or insect cell eukaryotic expression system to express the recombinant protein, and the expression product induces neutralizing anti-duck hepatitis A virus antibodies.

Methods for predicting cancer aggressiveness and / or risk of cancer recurrence in a subject are provided. The methods can include the steps of detecting histone gene expression at one or more replication-coupled histone genes in a biological sample from the subject and comparing the levels of histone gene expression at the one or more replication-coupled histone genes in the biological sample to a reference, wherein an increase in the levels of histone gene expression at the one or more replication-coupled histone genes in the biological sample compared to the reference is indicative of cancer aggressiveness and / or risk of cancer recurrence. Also disclosed are methods of ameliorating cancer aggressiveness and / or risk of cancer recurrence in a subject by reducing elevated histone gene expression at one or more replication-coupled histone genes in a subject.

This invention provides an application of gibberellin in the H3K27me3 modification of NAAT1 or HsfA2d and its role in regulating alkali-heat tolerance. It reveals that active gibberellin is associated with a series of genes, balancing the histone H3K27 trimethylation (H3K27me3) modification levels of NAAT1 and HsfA2d under alkali-heat stress, thereby enhancing the tolerance of gramineous plants to alkali-heat stress. This invention provides an important theoretical basis for breeding plant varieties or other plants with alkali-heat tolerance.

The invention discloses a method for regulating and controlling SLIT2 transcription and application of the method in ovarian granular cells, and belongs to the technical field of cell engineering and gene engineering. According to the invention, SLIT2e, SLIT2 gene promoter region histone acetylation and an SLIT2 gene are taken as research objects, and the SLIT2 gene promoter histone acetylation is verified through TSA or HDAC-siRNA to promote the expression of the SLIT2 gene promoter histone acetylation. The invention explores the mechanism that the enhancer regulates the gene promoter region histone acetylation and chromatin opening degree so as to regulate the transcription expression of the SLIT2 gene. The invention has a certain application value for researching the molecular mechanism of influence of histone acetylation of the SLIT2 gene promoter region on ovarian follicle development.