461 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 embodiment of the invention discloses a single cell transcriptome sequencing data annotation method. The method comprises the following steps: acquiring single cell transcriptome sequencing data associated with a target disease, and clustering a plurality of cells corresponding to the single cell transcriptome sequencing data to obtain a plurality of cell clusters; aiming at each marker gene corresponding to the target disease, according to single cell transcriptome sequencing data, obtaining the cell cluster gene expression quantity of the marker gene in each cell cluster; for each cell cluster and each cell type corresponding to the target disease, accumulating the cell cluster gene expression quantity of each marker gene corresponding to the cell type in all the marker genes in the cell cluster to obtain the total gene expression quantity; and aiming at each cell cluster, according to the total gene expression quantity corresponding to the cell cluster under each cell type, annotating a target cell type for the cell cluster from all the cell types. According to the technical scheme provided by the embodiment of the invention, the accuracy of cell type annotation can be ensured.

The invention discloses a mammalian cell expression vector, an expression plasmid and an expression system. The expression vector sequentially comprises the following elements: (a) a promoter; (b) a Kozak sequence; (c) a polyadenine sequence, the polyadenine sequence sequentially comprises a bGH PolyA sequence and a 5 'UTR PolyA sequence, the nucleotide sequence of the bGH PolyA sequence is as shown in SEQ ID NO.5, and the nucleotide sequence of the 5' UTR PolyA sequence is as shown in SEQ ID NO.6; (d) copying the start site; and (e) screening marker genes. According to the present invention, the expression vector is optimized by the expression element, such that the exogenous gene can be conveniently, efficiently and flexibly inserted and edited by the expression vector so as to achieve the stable extracellular or intracellular expression of the exogenous protein, and the expression efficiency of the exogenous protein can be improved by carrying out the expression plasmid construction based on the expression vector; and more efficient expression and purification of the foreign protein are 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 discloses regulation and control of a GmERECTAs gene (GmERs for short) on a soybean plant height and application of the GmERECTAs gene (GmERs for short). The invention belongs to the technical field of biology. An sgRNA sequence of a GmERs gene is recombined to a knockout vector pGES401 through a Golden Gate cloring technology. According to the method, four homologous genes of GmERs are knocked out by using a CRISPR / Cas9 technology, the four genes comprise GmERa, GmERb, GmERc and GmERd, and offspring screening is performed through herbicide resistance endowed by a Bar marker gene; after a positive result is obtained through screening, the GmERs gene knockout condition is determined through sequencing, and the plant height of a gene knockout soybean plant is remarkably reduced compared with that of a contrast. The invention has important significance for developing and breeding new soybean germplasm resources.

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 invention discloses a sebastes schlegeli testis cell line as well as a construction method and application thereof, and belongs to the technical field of marine fish cell culture. Primary culture and subculture are carried out on sebastes schlegeli juvenile fish testis tissues, so that the sebastes schlegeli testis cell line capable of being continuously subcultured in vitro for a long time is established. The sebastes schlegeli testis cell line established by the invention is in a fibroblast shape, has a good growth curve, and has relatively strong multiplication capacity. The lentiviral vector can significantly express the stromal cell marker gene temp2, can transfect the cell line, and can be subsequently applied to research of gonad development related functional genes. At present, the cell line has been subjected to in-vitro continuous passage for more than 60 generations, can be subjected to cryopreservation, and is high in recovery survival rate. Establishment of the cell line provides a stable and excellent experimental material for research of fish gonad development genes.

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.

A method for inferring super-resolution tissue architecture by integrating spatial transcriptomics with histology includes receiving at least one histology image of a tissue sample at a histology feature extractor. The histology feature extractor partitions the at least one histology image into image tiles and partitions each of the image tiles into image sub-tiles. The histology feature extractor extracts histology features from the histology image, the extracted histology features include low-level image features extracted from the image sub-tiles and high-level image features extracted from the image tiles. A super-resolution gene expression predictor predicts gene expression for the image sub-tiles using the extracted histology features and a predictor model trained with spot-level gene expression observations. A tissue architecture annotator clusters the image sub-tiles based on the predicted gene expression of the image sub-tiles and annotates each of the image sub-tiles using the predicted gene expressions and a marker gene reference panel.

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 discloses a site-specific integration site screening method, a carrier particle, a site-specific integration method and a particle. The invention provides a site-specific integration site screening method, a carrier particle, a site-specific integration method and a particle, and aims to integrate upstream and downstream marker genes with recognition sequences on the carrier particle, perform protein expression quantity screening while performing monoclonal screening, obtain a monoclonal carrier with good performance through one-step screening, and perform site-specific integration on the carrier particle. The marker gene is replaced when the exogenous target gene is expressed, so that the overall stability of the particle is greatly ensured, the overall expression performance and stability of the expression particle are ensured while the development of the expression particle is shortened, and the technical problem that an expression vector in the prior art is relatively long in development time or poor in performance is solved. The site-specific integration site screening method provided by the invention can effectively shorten the development period of the expression vector with high expression quantity and overall stability. The carrier particle provided by the invention is good in adaptability and good in expression quantity and passage stability.

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 a method and a system for intelligently analyzing heterogeneity of endothelial cells in atherosclerosis. The method comprises the following steps: acquiring sample data of the endothelial cells in the atherosclerosis of a patient; performing feature extraction on the sample data of the endothelial cells to obtain gene expression features of the endothelial cells; dividing the endothelial cells into different subgroups according to gene expression characteristics of the endothelial cells and gene expression similarity so as to determine marker genes for distinguishing the different subgroups, and defining and representing characteristics of each subgroup by analyzing expression modes of the marker genes; gene expression differences among different endothelial cell subgroups are compared, differential expression genes of the related endothelial cell subgroups, which are significantly up-regulated or down-regulated in the atherosclerosis process, are identified, and the differential expression genes are used as key molecular markers for understanding functional changes and heterogeneity of endothelial cells. According to the method, the heterogeneity of the endothelial cells in the atherosclerosis can be accurately analyzed.

The invention belongs to the field of rice cultivation, and particularly relates to a group of marker genes for identifying a rice grain filling process and application thereof. The marker gene screened by the invention is expressed in a monotonically increasing or decreasing manner during the rice grain filling period, and can be used as the marker gene to accurately reflect the time progress of rice grain filling. By detecting the expression level of the marker genes, the rice grain filling process can be identified in a molecular manner, and the duration of the rice grain filling process can be determined.

The invention relates to the technical field of biology, in particular to a benzene-induced blood abnormality marker, an evaluation model and a construction method and application of the benzene-induced blood abnormality marker and the evaluation model, and provides the following scheme that the method comprises the steps that S1, principal component analysis is conducted on benzene damage expression profile data sets, then preliminary screening conditions are set, and differential expression genes between the benzene damage expression profile data sets are screened out; s2, carrying out GO, KEGG and GSEA enrichment analysis on the differentially expressed gene to obtain an analysis result; s3, different machine learning algorithms are used for screening characteristic genes from analysis results, intersection is taken, key genes are obtained, then verification analysis is conducted on the key genes, and benzene damage marker genes are obtained; and S4, training and testing a plurality of learning models by using the benzene injury marker gene, and selecting a proper learning model as a benzene-induced blood abnormality evaluation model according to a test result of the learning model. According to the invention, a theoretical basis can be provided for early intervention of benzene toxicity, and a more comprehensive and sensitive model can be provided for risk assessment.

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 relates to an expression and methylation quantitative detection method of a ZNF300P1 gene and application of the ZNF300P1 gene in assisting myeloid tumor diagnosis and prognosis evaluation. The invention provides ZNF300P1 gene expression and methylation quantitative detection, and the ZNF300P1 biomolecular marker gene is shown as SEQ ID NO.1. According to the expression and methylation detection primer provided by the invention, the ZNF300P1 biomolecular marker expression detection primer is shown as SEQ ID NO.2-3, the ZNF300P1 biomolecular marker methylation detection primer is shown as SEQ ID NO.6-9, and the ZNF300P1 biomolecular marker methylation detection primer is shown as SEQ ID NO.2-3. The kit is used for detecting the expression and methylation level of bone marrow / peripheral blood ZNF300P1 of patients with myelodysplastic syndromes and acute myelogenous leukemia, serves as a novel biomarker of the patients with myelodysplastic syndromes and acute myelogenous leukemia, is combined with other indexes, and provides support for early diagnosis and prognosis evaluation of the patients.

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.

Methods are provided for amplifying microbial cell free DNA (mcfDNA) in blood samples. In contrast to amplicon sequencing, in which regions between two conserved sequences are targeted for PCR amplification, the methods herein use a single conserved sequence as the primer annealing site to initiate PCR amplification. Enrichment PCR (E- PCR) is performed using a degenerate primer complementary to a conserved region in a microbial phylogenetic marker gene and an additional primer to the opposite end of the mcfDNA. The conserved region is adjacent to a hypervariable region. Amplification PCR is performed on the enriched mcfDNA fragments using a modified degenerate primer having a 3 '-end extension relative to the degenerate primer, and the additional primer, thereby generating enriched amplified mcfDNA fragments. Sequencing the amplified mcfDNA enables identification of microbial species, which is useful for diagnosing and monitoring inflammatory diseases and cancer, including stratification of cancer by stage, type, and tumor subtype.

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.

Provided are a method for screening a site-directed integration site, a vector microparticle, a site-directed integration method and a microparticle. Provided are a method for screening a site-directed integration site, a vector microparticle, a site-directed integration method and a microparticle. The objective of the present invention lies in that by integrating a marker gene having recognition sequences upstream and downstream on a vector microparticle, protein expression screening and single clone screening are simultaneously performed so as to obtain a single clone vector having good performance by means of one-step screening, and the marker gene is then replaced when an exogenous gene of interest is expressed so as to greatly ensure the overall stability of the microparticle, which ensure the overall expression performance and stability of the expression microparticle while shortening the development of the expression microparticle, thereby solving the technical problem in the prior art that expression vectors are long in development time or poor in performance. The provided method for screening a site-directed integration site can effectively shorten the development period of expression vectors having high expression and overall stability. The provided vector microparticle has good adaptability, good expression and good passaging stability.

The invention discloses a soybean genetic transformation method for improving positive transformation rate, which comprises the following steps of: infecting a soybean explant with a recombinant expression vector containing a target gene and a marker gene through agrobacterium liquid; an ALS inhibitor is added into a bud induction culture medium and a bud elongation culture medium to improve screening culture of the positive conversion rate of the explants, and a transformant is obtained. According to the soybean genetic transformation method which takes the gene cloned from the cynodon dactylon as the marker gene and the ALS inhibitor as the screening agent, the positive transformation rate is increased to 6% or above, the maximum is 12.05%, and the soybean transformation efficiency is remarkably improved.