359 results about "Marker gene" patented technology

The invention relates to a fluorescent protein selection marker staygold-levEr gene as well as a construction method and application thereof, and belongs to the technical field of biological engineering. The invention provides a fluorescent protein selection marker staygold-levEr gene for screening an L-valine production strain and a construction method of a gene fragment, compared with other similar gene selection markers, the selection marker provided by the invention has the advantages that the fluorescence intensity is positively correlated with the concentration of L-valine, and the screening marker has the advantages that the screening marker can be used for screening the L-valine production strain; according to the screening marker, the screening efficiency of the L-valine high-yield strain can be obviously improved, the screening time and cost can be saved, the screening efficiency can be improved, and the screening marker has a wide application prospect.

The invention belongs to the technical field of biology, and discloses a reagent combination or kit for constructing intestinal organs and application of the reagent combination or kit. According to the reagent combination or the kit, intestinal organs can be obtained from cell-derived epithelial cells obtained from a donor in a non-invasive manner, and the obtained intestinal organs can be cryopreserved and recovered and can be amplified in vitro for a long time; transcriptome characteristics are similar to those of real human small intestine tissues, and typical marker genes of various small intestine pedigree cell types are highly expressed; compared with intestinal organs obtained through induction of pluripotent stem cells, the intestinal organs have more intestinal pedigree characteristics, and the intestinal function development is more mature; after being promoted to be mature, the intestinal organ also highly expresses genes related to drug absorption and metabolism, has drug absorption capability similar to that of an immortalized intestinal cell line, but more prominently shows intestinal cell lineage characteristics, is closer to an intestinal environment in a real human body, and can be used for screening intestinal disease drugs; the intestinal barrier function is realized.

The invention provides a fluorescent protein selection marker staygold-ygjHr gene and application thereof, and belongs to the technical field of biological engineering. The preparation method comprises the following steps: replacing codons of L-valine in nucleotide sequences of a fluorescent protein gene Staygold and a gene ygjH by using a rare codon GTC to respectively obtain a fluorescent protein gene Staygoldr after codon replacement and a gene ygjHr after codon replacement, and connecting the two segments of genes after codon replacement by using a flexible protein peptide to obtain the fluorescent protein selection marker Staygold-ygjHr gene. Experiments prove that the gene can obviously improve the screening efficiency of L-valine high-yield strains.

The invention provides an enhancer RNA (Ribonucleic Acid) molecule MZGAe1 and application thereof, and relates to the technical field of biology. The invention provides an enhancer RNA (Ribonucleic Acid) molecule MZGAe1. The nucleotide sequence of the enhancer RNA molecule MZGAe1 is as shown in SEQ ID NO: 1; meanwhile, the invention further provides a specific sgRNA sequence for activating the molecule in a targeted manner and a recombinant vector of the specific sgRNA sequence. Expression of endogenous MZGAe1 of cells is specifically activated by adopting a CRISPR activation technology, and conversion of mouse embryonic stem cells to bicellular-like cells can be efficiently promoted. By providing a brand-new endogenous targeted MZGAe1 accurate activation tool, the efficient transformation of wild mouse embryonic stem cells to bicellular cells can be realized only under the condition of endogenous activation of single enhancer RNA molecule MZGAe1, the proportion is at least increased by 5%, the expression of a totipotent marker gene ZSCAN4 is activated, and the expression of the totipotent marker gene ZSCAN4 is promoted. And an efficient and specific brand-new research tool and scheme are provided for researching zygote genome activation, embryonic development early events and cell reprogramming.

The present invention relates to the field of tissue-derived cell identification, in particular to a method for identifying tissue-derived cells in the body fluid based on single-cell sequencing technology. In the present invention, on the basis of high-throughput single-cell RNA sequencing data of cell samples obtained from body fluids, by means of reference component analysis (RCA), expression of known tissue-related marker genes, and a tissue-derived cell prediction model constructed based on logistic regression, the specific identification of tissue-derived cells in body fluids is achieved.

The invention provides a fluorescent protein selection marker staygold-ynaMr gene and application thereof, and belongs to the technical field of biological engineering. The method comprises the following steps: replacing codons of L-isoleucine in nucleotide sequences of a fluorescent protein gene staygold and a gene ynaM by using a rare codon ATA to respectively obtain the fluorescent protein gene staygold after codon replacement and the gene ynaMr after codon replacement, and connecting the two segments of genes after codon replacement by using a flexible protein peptide, and the fluorescent protein selection marker staygold-ynaMr gene is obtained. Experiments prove that the gene can obviously improve the screening efficiency of L-isoleucine high-yield strains.

The invention discloses a key marker gene-based intelligent cell type annotation method, which comprises the following steps of: constructing a static knowledge base by using known marker genes in a reference database, and endowing the marker genes with cell specific weights by using a TF-IDF method, so that the annotation accuracy and interpretability are improved. Meanwhile, under the condition that static matching is insufficient, the literature is understood through a large language model, mark information is extracted, dynamic completion of the knowledge base is achieved, the defect that updating of a traditional knowledge base is lagged is overcome, and good adaptability and expansibility are achieved. Besides, static and dynamic matching scores are fused in the annotation process, so that more robust cell type identification is realized, annotation requirements of multi-tissue, multi-species and novel cell states are adapted, high-precision and extensible cell type annotation can be realized in a scene with insufficient reference knowledge or a fuzzy sample, and the annotation efficiency is improved. And the method has good universality and practicability.

The invention belongs to the technical field of biotechnology and agricultural biological breeding, and relates to a sorghum double haploid breeding method which comprises the following steps: (1) performing gene mutation on SbMTL and SbDMP to obtain a sorghum double-gene mutant strain according to any one of claims 1 to 3; (2) screening and identifying sorghum haploids by using GFP (Green Fluorescent Protein) marker and Ruby marker genes; and (3) naturally doubling the obtained haploid plant to obtain the double haploid of sorghum. SbMTL and SbDMP genes of sorghum are knocked out by utilizing a CRISPR / Cas9 technology so as to realize high-frequency haploid induction, meanwhile, an effective haploid selection marker is introduced, the induction frequency of a haploid induction system is evaluated, and after the obtained haploid plant is naturally doubled, a double-haploid system of sorghum is successfully obtained. The induction efficiency of the sorghum haploid induction line reaches up to 18% or above, the economic value is high, and the process of sorghum breeding is remarkably promoted.

The invention discloses an oat KASP marker, a fingerprint spectrum and application, and relates to the technical field of molecular breeding. The KASP marker comprises 96 SNP (Single Nucleotide Polymorphism) sites, and the position information and the polymorphism of the KASP marker are shown in a table 1. The invention also provides a gene chip for analyzing the KASP marker and an oat variety characteristic fingerprint spectrum constructed by using the KASP marker, and the KASP marker, the gene chip and the oat variety characteristic fingerprint spectrum provided by the invention can be used for screening, identifying and tracing oat quality. The method can be used for identifying the authenticity of the seeds and the genetic relationship between the parent and offspring of the oat, and has important significance for protecting and improving the germplasm resources of the oat.

The invention provides a cell type annotation method and device based on plant single cell transcriptome data and a readable storage medium. Annotation is achieved through multi-level data integration, wherein preliminary annotation is expressed based on cell type marker genes or homologous genes; calculating expression correlation auxiliary annotations of the to-be-analyzed data and the known transcriptome data set; performing function enrichment on the cell cluster differential genes to deduce cell types; carrying out quasi-timing analysis on the heterogeneous cell clusters and annotating subgroups; and finally integrating and generating a comprehensive annotation. The method solves the problem that the prior art depends on artificial experience and is insufficient in basic data set, and is suitable for mode and non-mode plants.

The invention discloses a construction method of a trophoblast organ-like model, the trophoblast organ-like model and application thereof, and relates to the technical field of cell model.The construction method comprises the steps that placenta villus tissue is aseptically separated, gradient separation and purification are conducted to obtain extraembryonic early cells, the extraembryonic early cells are mixed with matrigel, and the trophoblast organ-like model is obtained; and carrying out 3D culture by using a culture medium containing early-stage and later-stage differentiation compositions to obtain the organ-like model with various trophoblast cell lineages. Based on the model, placenta toxicity research is carried out on pregnancy drugs, and the research specifically comprises the degree of 3D organoid form destruction after drug treatment, organoid activity and the expression condition of trophoblast marker genes. Compared with a traditional two-dimensional cell model, the placental toxicity detection method has the advantages that the placental toxicity of the medicine is detected by taking the trophoblast-like organ as the model, the accuracy is higher, the predictability is higher, and the influence of the medicine on human placental tissues is accurately reflected.

The invention discloses application of an overexpressed carp lpla gene in regulation and control of expression of cyprinid fish tissue-related genes, and relates to the field of gene engineering, in particular to application of the overexpressed carp lpla gene in regulation and control of cyprinid fish tissue-related genes. The overexpressed carp lpla gene is applied to regulation and control of contents of lipid indexes of total cholesterol TC and triglyceride TG of different tissues of cyprinid fishes. The invention further discloses application of the overexpressed carp lpla gene in regulation and control of fat marker genes lpla, ppar gamma, Fabp3, fast and acaca of livers and muscle tissues of cyprinid fishes. The invention further discloses application of the overexpressed carp lpla gene in regulating and controlling the relative expression quantity of carp liver cell fat metabolism related genes lpla, ppar gamma, Fabp3, fast, acaca, hsl and pnpla2. The overexpressed carp lpla gene can regulate and control lipid deposition in different tissues in zebra fish, and a research model is provided for lpla to regulate and control fat deposition and differentiation. According to the overexpressed carp lpla gene disclosed by the invention, lpla high-expression carp hepatocytes are obtained through a liposome transfection technology, so that fat key gene expression in the carp hepatocytes is changed.

The present invention belongs to the field of bioengineering, genetic engineering technology and oligosaccharide (rare sugar) synthesis technology, and specifically relates to a method for producing lactose-N-disaccharide using genetically modified recombinant yeast. The technical scheme of the present invention is: a recombinant yeast cell is introduced with (1) a heterologous lactose-N-disaccharide phosphorylase ( LNBP ) gene, and (2) fructose-1-phosphate phosphorylase ( YQAB ) gene, and further preferably, also includes (3) destruction or modification of a marker gene. The recombinant yeast cell has the ability to synthesize lactose-N-biose (Lacto-N-biose, LNB) from lactose, and the yield of LNB is greatly improved by a molecular switch.

The invention discloses application of a KASP molecular marker in identification or auxiliary identification of soybean hundred-grain weight character, and belongs to the technical field of soybean agronomic character molecular marker development and molecular marker auxiliary breeding. According to the application of the KASP molecular marker in identification or auxiliary identification of the soybean hundred-grain weight character, the genotype of the KASP molecular marker is G or A, and the KASP molecular marker is located at the 7703700bp position of the No.6 chromosome of a soybean genome Wm82. A2. V1. The obtained KASP marker for regulating and controlling the hundred-grain weight character gene of the soybean is suitable for rapid screening and molecular marker-assisted breeding of the relative height of the hundred-grain weight of the soybean. The molecular marker can detect genotypes of materials with different hundred-grain weights in natural populations, can also be used for molecular-assisted selective breeding of soybean materials with different hundred-grain weights, replaces large-scale phenotypic screening, reduces field workload, and greatly improves screening accuracy and efficiency.

The invention discloses a structural variation molecular marker located on a pig chromosome 7 and related to the lactation ability of a sow. The structural variation molecular marker is a DNA (deoxyribonucleic acid) fragment which is inserted between the 112024896 bp site and the 112024896 bp site on a chromosome 7 of an international pig reference genome 11.1 version, wherein the nucleotide sequence of the DNA fragment is shown as SEQ ID NO: 1; the genotype of the gene is T / T, T / INS and INS / INS. The structural variation molecular marker provided by the invention obviously influences the total weight gain character and the lactation ability character of the 7-day-old sow, and by breeding the pig with the structural variation molecular marker genotype of T / T, the lactation ability of the sow can be obviously improved, and the survival rate and the weaning weight of piglets are increased.

The invention relates to the technical field of biological detection, and discloses a reagent composition and a kit for detecting cervical cancer and precancerous lesions of cervical cancer and application of the reagent composition and the kit. The reagent composition provided by the invention is used for detecting the methylation degree of a specific marker gene, and the detection sensitivity and specificity of cervical cancer and high-grade cervical cancer precancerous lesions are at a relatively high level. When the reagent composition is used for screening cervical cancer and precancerous lesions of HPV positive people, compared with conventional cytological detection, the colposcope referral rate is greatly reduced, meanwhile, the detection times for definite diagnosis of high-grade cervical lesions or cervical cancer are reduced, and the waste of medical resources and the examination burden of patients are reduced.

The invention discloses research of Pdh1 allelic variation in soybean breeding material distribution and pod cracking resistance, and pod cracking severely affects improvement of soybean productivity and is one of important indexes of soybean breeding selection. The invention discloses a high-quality anti-pod-cracking soybean breeding identification marker Pdh1-2 gene, the Pdh1-2 gene is a variation type of a Pdh1 gene, the base sequence of the Pdh1 gene is as shown in SEQ ID NO.2, and the base sequence of the Pdh1-2 gene is as shown in SEQ ID NO.3. The invention also discloses a preparation method of the high-quality anti-pod-cracking soybean breeding identification marker Pdh1-2 gene. The practicability of the Pdh1-2 variation type is further explored. 1118 breeding strains are randomly selected, the proportions of split pods and non-split pods of variation types are respectively counted, and the result shows that 975 parts of materials are Pdh1-1 and Pdh1-3 variation types, and 143 parts of materials are Pdh1-2 variation types. The combined phenotypic analysis shows that the proportion of the Pdh1-2 split pod material is the maximum and is 84.6%, the proportion of the Pdh1-1 is 81.0%, and the proportion of the Pdh1-3 is the minimum and is only 41.1%. The research provides a theoretical basis for breeding of southern anti-pod-cracking soybeans assisted by the molecular marker.

The invention belongs to the field of tilapia mossambica breeding, and discloses a rapid and efficient tilapia mossambica breeding method and system.The method comprises the steps that tilapia mossambica populations with excellent character potential are collected and screened, and a basic breeding population is established; molecular markers related to target characters are developed, and individuals carrying excellent genes are rapidly screened; a target gene is precisely regulated and controlled through a CRISPR / Cas9 technology, so that stable inheritance of excellent characters is accelerated; phenotypes such as growth performance, disease resistance and environment resistance are efficiently evaluated and screened; the excellent characters are fixed; and the performance of the new strain under the actual breeding condition is evaluated. The efficient and low-cost tilapia mossambica breeding method is constructed by integrating molecular marker, gene editing and phenotype screening technologies, the breeding efficiency and the commercialization value of a new strain are remarkably improved, and powerful technical support is provided for the aquaculture industry.

The invention discloses a method for screening a biomarker for early non-invasive Alzheimer's disease screening based on machine learning. The method comprises the following steps: acquiring blood-derived cell free RNA (Ribonucleic Acid) sequencing (cfRNA-seq) and brain-derived single cell transcriptome sequencing (scRNA-seq) data of an Alzheimer's disease (AD) patient and an age-matched contrast; and based on the standardized data, screening out genes presenting the same expression mode in the two types of data as the biomarker for non-invasive Alzheimer's disease screening. According to the method, 34 characteristic genes which are jointly and differentially expressed in cfRNA and scRNA data sets are found, and the AD diagnosis classifier is successfully constructed by using the 34 characteristic genes. A diagnosis classifier can accurately predict AD patients and effectively distinguish AD early-stage patients, and meanwhile, the risk of an individual suffering from AD is evaluated. Results show that the key marker genes can be applied to early AD non-invasive screening and prevent disease progression; meanwhile, AD patients with different disease progresses can be distinguished, and support is provided for personalized treatment schemes of the AD patients.

The invention discloses application of a compound with a benzo-aza structure in preparation of a medicine for inhibiting osteoclast formation, the structure of the compound is shown as a formula (1), and R substituent is one of H, OCH3, Br, F and i-Pr. Cell experiments prove that the compound with the benzo-aza structure has a remarkable inhibiting effect on RANKL-induced osteoclast differentiation and marker gene expression, can be used as a candidate medicine which is novel in structure and is used for inhibiting osteoclast formation, and is used for targeting osteoclast to treat osteoporosis related diseases.

The invention discloses a primer composition and a kit for identifying testudinate species and application of the primer composition and the kit. The primer composition comprises primers (SEQ ID NO.1-6) for targeted amplification of specific marker genes 16S, COI and Cytb of testudinate mitochondrial species and an enrichment universal primer (SEQ ID NO.7). According to the invention, full lengths of three mitochondrial genes 16S, COI and Cytb are taken as targets, a specific primer group added with an enrichment tag and an enrichment primer are designed, and a targeted nanopore sequencing method capable of realizing testudinate species identification is established and optimized, and the method can simultaneously capture testudinate mitochondrial species specific marker gene information of a family species level, and can be used for identifying testudinate species. The problem that the existing DNA bar code is difficult to accurately identify the testudinate species due to the lack of database information is effectively solved; and each marker gene can obtain a complete coding sequence, so that genetic information is richer, and the method can be used for geographical traceability analysis of testudinate species.

The present invention discloses a method for establishing and applying a caudal fin-induced pluripotent stem cell line of Erythroculter ilishaeformis. By the tissue adherence method, a cell line with high proliferation performance and stemness is isolated from the caudal fin of Erythroculter ilishaeformis. In the early stage, the cells of this cell line are significantly fibroblast-like. Later, the cells become smaller but still maintain a spindle shape, without obvious contact inhibition phenomenon, and can spontaneously form cell clusters similar to induced pluripotent stem cells. Alkaline phosphatase staining is positive and it expresses stemness marker genes such as Oct4, Sox2, Klf4, Nanog, and c-Myc. The present invention uses a single cAMP agonist small molecule compound to reverse the phenomenon of weakened stemness after continuous subculture of this cell line, enabling it to maintain the characteristics of high proliferation performance and high stemness, which is suitable for future research on the induction and differentiation of fish stem cells and provides more choices for the research and application of fish stem cells.

The invention discloses a brain glioma animal model construction method based on an AAV mediated immune microenvironment. The method comprises the following steps: selecting an immunodeficient mouse at the right age; human CD34 + hematopoietic stem cells are injected for immune system reconstruction; human CD45 + immune cells in the peripheral blood PBMC are detected; the method comprises the following steps: selecting tumor tissues with high-expression brain glioma marker genes, carrying out rapid qPCR (quantitative polymerase chain reaction) or targeted RNA-seq immunohistochemical multiple verification, and screening consistent tumor tissues; the method comprises the following steps: physically dicing tumor tissues, treating the tumor tissues with pretreatment liquid, digesting the tumor tissues into single-cell suspension, and inoculating the single-cell suspension into a mouse cranium; an AAV vector carrying a specific immune regulation gene is injected into the tail vein of the mouse, and the brain glioma immune microenvironment is regulated; when it is observed that the tumor tissue grows stably and has immune cell infiltration characteristics, construction of the brain glioma animal model is completed. Compared with an existing model, the brain glioma model which is more stable, more scientific and closer to the human immune background is provided, and the brain glioma model is suitable for related immunotherapy drug screening and mechanism research.

The present application relates to the field of agricultural biotechnology, and specifically discloses a kind of corn ear roughness related molecular marker and its application.The present application provides the application of corn ear roughness molecular marker in ear roughness corn molecular marker assisted breeding, the molecular marker is Zm00001d020000 gene, the nucleotide sequence of the Zm00001d020000 gene is as shown in SEQ ID NO:1, the expression amount of the gene is positively correlated with the corn ear roughness trait, multiple SNP sites related to ear roughness are screened out by GWAS analysis, and simultaneously, QTL point analysis is used to be commonly positioned to SNP_84054216 located on the 7th chromosome, and the marker gene Zm00001d020000 for regulating ear roughness is excavated from the site, and the results of the present application provide technical support for molecular marker breeding of rough ear corn varieties.

The present application relates to the field of genetic engineering, and in particular to a vector for analyzing plant promoter expression specificity, a preparation method and application thereof. The T-DNA region of the vector comprises, from 5' end to 3' end, a callus-specific promoter-driven selection marker gene expression cassette and a reporter gene expression cassette with a multiple cloning site; the callus-specific promoter-driven selection marker gene expression cassette comprises a callus-specific promoter, a selection marker gene coding sequence and a terminator. The vector of the present application utilizes a rice callus-specific promoter to drive the expression of a selection marker gene, can greatly reduce the non-specific interaction with the promoter of a target gene, improve the specificity of the expression of the target gene, and effectively reduce the biological safety risk caused by the selection marker gene in a transgenic plant, and has important application value in basic theoretical research and molecular breeding.

The invention belongs to the technical field of biomolecular markers, and particularly relates to a cross-animal general skeleton-derived hematopoietic stem cell marker gene set and a screening method thereof. According to the invention, a bone-derived hematopoietic stem cell marker gene set universal across animal categories is constructed for the first time, and the bone-derived hematopoietic stem cell marker gene set comprises at least five of Cdc42, Cbx, Tfam, Denr, Mcts1, Ak2, Ruvbl, Ahcy, Nna and Vdac; the hematopoietic stem cell marker gene set is obtained through cross-species homologous gene screening, the defect that a traditional vertebrate marker has no orthohomology in invertebrates is overcome, accurate recognition of HSC in shells is achieved, the hematopoietic stem cell marker gene set has species universality and cell specificity, the immune state of aquatic animals can be evaluated, disease-resistant breeding can be guided, and the application prospect is wide. And molecular evidence is provided for analyzing an evolution path of a hematopoietic system from invertebrates to vertebrates.

The invention discloses application of a PRTFDC1 gene in promoting porcine skeletal muscle satellite cell differentiation, myotube formation and skeletal muscle development. According to the invention, overexpression and knock-down experiments of the PRTFDC1 are respectively carried out in pig MuSCs, and immunofluorescence, qRT-PCR and Western blot analysis are carried out on cells subjected to induced differentiation, so that the PRTFDC1 is effectively shown to play a positive regulation role in a pig MuSCs differentiation process, and a new marker gene which can be applied to pig molecular breeding and growth performance evaluation is provided; and a novel molecular target and a novel detection tool are provided for muscle development regulation and biological breeding.

The invention relates to the technical field of biology, in particular to a gene co-expression network-based sepsis marker screening method and application. The screening method comprises the following steps: screening differential expression genes; performing hierarchical clustering analysis on each differential expression gene to combine each differential expression gene into a plurality of target gene modules; and screening out hub genes from each target gene module. According to the method, gene expression data and clinical feature data are combined, the accuracy of screening sepsis marker genes is improved, possible abnormal genes irrelevant to the pathophysiological process of sepsis are avoided, gene modules are divided more finely through primary clustering analysis and secondary clustering analysis, GO function analysis is combined, and the accuracy of screening the sepsis marker genes is improved. According to an analysis result, selection of a GO term subset is optimized, and a step length set by primary clustering is corrected, so that the fine granularity and accuracy of analysis are improved, division of gene modules is optimized, and more refined treatment and screening of markers of sepsis are realized.