333 results about "Multi gene" patented technology

According to the invention, the primers are designed for 65 leukemia fusion genes through sufficient literature investigation in the early stage, the leukemia genotypes covering 90% or more are covered, and the primers are good in amplification performance and strong in specificity; based on a multiple digital PCR system of an innovative multicolor fluorescent channel, different fluorescently-labeled hydrolysis probes are designed, multi-gene multi-channel detection is combined, 65 leukemia fusion genes are detected at the same time, leukemia typing can be judged through one-time detection, and time is saved. Through high-throughput detection, the cost is reduced to 1 / 10 of that of foreign like products, and the sensitivity is high.

The invention belongs to the technical field of molecular genetic breeding, and particularly relates to a molecular marker Zm00001eb181850 gene related to corn leukoderma resistance and application thereof, the invention provides a molecular marker related to the corn leukoderma resistance, the gene sequence of the molecular marker is shown as SEQ ID NO: 1 and / or SEQ ID NO: 2, and the molecular marker Zm00001eb181850 gene is shown as SEQ ID NO: 1 and / or SEQ ID NO: 2. Researches prove that corn white spot resistance QTL qMWS4-1 and qMWS4-5 intervals and resistance SNP: 4-118983592 have co-localization, the corn white spot resistance QTL has a high phenotypic interpretation rate, the associated gene Zm00001eb181850 (SEQ ID NO: 2) is highly expressed in corn leaves, it is proved that the associated gene Zm00001eb181850 (SEQ ID NO: 2) is associated with the corn leaf spot, the corn white spot resistance is a quantitative character jointly regulated by multiple genes, and the corn leaf spot resistance QTL has a good application prospect. According to the technical scheme provided by the invention, a new method is provided for cultivating a new variety of leukoderma-resistant corn by utilizing a modern molecular breeding technology.

The invention relates to the technical field of high-throughput sequencing, and discloses a kit and a library construction method and a target variation detection and analysis method thereof, the kit comprises a pre-library construction reagent and a target region liquid phase hybridization capture reagent; wherein the pre-library construction reagent comprises a repair reagent, a joint, a connection reagent, an amplification reagent and purified magnetic beads; the target area liquid phase hybridization capture reagent comprises a hybridization reagent, a cleaning reagent, an amplification reagent, a purification magnetic bead, a target gene probe and a CNV / LOH probe. The invention provides a rapid, accurate and low-cost construction method of a library containing a targeting gene probe and a CNV / LOH probe and application of the library, through one-time library construction and sequencing, the advantages of a high-throughput targeting sequencing technology and a CNV-Seq technology are integrated; multi-gene point mutation, insertion and deletion, gene copy number variation, large-fragment copy number variation and heterozygosity deletion can be detected at the same time.

The invention provides a primer probe composition and a kit for detecting a Databean virus and application, and relates to the technical field of biological detection, the primer probe composition comprises a primer probe group aiming at a Databean virus S gene and a primer probe group aiming at a Databean virus L gene, the Databean virus L and S genes are used as amplification target genes for design, and the kit is used for detecting the Databean virus. The primer probe group aiming at the S gene of the Databu virus and the primer probe group aiming at the L gene of the Databu virus are used as the primer probe composition, so that multi-gene target amplification is realized, and the positive detection rate reaches 100% and is higher than the positive detection rate of single-gene amplification; the lowest detection limit is up to 200 copies / mL; the quantification limit reaches 500 copies / mL, and the quantification linear range is 5.0 * 10 < 2 >-1.0 * 10 < 8 > copies / mL. The technical problem that in the prior art, a high-sensitivity and high-accuracy multiplex qPCR detection method for the nucleic acid of the copyda virus is lacked is solved.

The present invention relates to a method for constructing an eight-gene-edited xenogeneic organ transplantation donor pig, belonging to the field of animal biotechnology. In the wild-type porcine fetal fibroblast cell line, the GGTA1, β4GalNT2, and CMAH genes are knocked out by using the CRISPR / Cas9 gene editing technology, and the humanized genes of hCD39, hCD46, hCD55, hCD59, and hTBM are transfected. Combining with somatic cell cloning technology, GTKO / β4GalNT2KO / CMAHKO / hCD39 / hCD46 / hCD55 / hCD59 / hTBM eight-gene-edited cloned pigs are constructed. Further, through genotype, mRNA, protein expression identification and functional analysis, eight-gene-edited xenogeneic organ transplantation donor pigs are obtained. The present invention solves the technical problems of high production difficulty, low efficiency, and low survival rate of donor pigs for multi-gene-edited xenogeneic organ transplantation, and maximally solves the common problems of immune rejection reaction and complement dysregulation faced during xenogeneic organ transplantation, laying a foundation for more targeted development of donor pigs suitable for different tissue and organ xenotransplantation, and having important value for promoting the clinical transformation of xenogeneic organ transplantation.

The invention discloses a rhodosporidium toruloides strain with high homologous recombination efficiency as well as a construction method and application thereof, and belongs to the field of oil-producing yeast engineering bacteria. According to the rhodosporidium toruloides strain, the rhodosporidium toruloides strain NP11-SaCas9-1 is used as an original strain, and an RAD52 gene is over-expressed. The modified yeast engineering bacterium provided by the invention can realize high homologous recombination through CRISPR system mediation, the homologous recombination efficiency reaches 90-100%, a multi-gene editing platform can be constructed on the basis, and the modified yeast engineering bacterium can be applied to the fields of synthetic biology, metabolic engineering, genetic engineering and the like, so that the modified yeast engineering bacterium is simpler, quicker, more convenient and more accurate in the genetic modification process. And the homologous recombination activity of the rhodosporidium toruloides can be improved by expressing the homologous recombination related protein Rad52, and the genetic engineering strategy can be used for modifying other unconventional yeasts.

The invention provides a polygene base editing carrier, an editing method and application, and belongs to the technical field of gene editing. According to the polygene base editing vector provided by the invention, the mode that the replicon element is connected with U6 and sgRNA in series is utilized, so that the plasmid containing the replicon element can continuously exist in a cell, and other elements on the plasmid can be continuously expressed in the cell, so that the polygene silencing efficiency is improved. The invention also provides a tandem carrier, namely an epi element in the carrier is removed. The two multi-gene editing vectors constructed in the embodiment of the invention have silence editing effects on targeting sites, and the editing efficiency of the vector containing the epi element is higher. According to the invention, a 5-gene silent homozygous cell line is obtained through a drug enrichment screening method, and a material and a method are provided for disease-resistant breeding.

The invention relates to a polygene methylation detection kit for liver cancer, a detection system and application thereof, and aims to provide a marker combination for early liver cancer screening. The kit is used for detecting the methylation state of one or more of 114 genes of BDH1, CPAMD8, DAB2IP, DLEC1, HOXA1, MIXL1, OTX1, PCDH8, RNF135, SIX2, SPINT2, TACSTD2, VIM and ZAR1, and is combined with restriction enzyme digestion and fluorescent quantitative PCR (Polymerase Chain Reaction) technologies, so that the early diagnosis of the liver cancer is realized. The kit and the system provided by the invention have high accuracy, specificity and early diagnosis capability, and experiments prove that the detection method can effectively distinguish liver cancer tissues from para-carcinoma tissues, and has relatively high clinical application value. The method is not only suitable for early screening of liver cancer, but also can provide a scientific basis for early treatment of liver cancer patients.

The present invention relates to a method for the ex vivo molecular diagnosis of gastrointestinal stromal tumors, comprising the steps of: providing at least one isolated tissue sample comprising at least one nucleic acid and the subsequent extraction of the at least one nucleic acid. The method according to the present invention further comprises a step of quantifying the at least one previously isolated nucleic acid. Furthermore, the method according to the present invention comprises preparing at least one amplicon library of said previously isolated nucleic acid, which comprises sequences obtained from the FGFR1, NF1, SDHA, SDHB, SDHC and SDHD genes. This step is followed by the step of purifying the at least one amplicon library and the step of simultaneously sequencing the amplicons themselves. The present invention also relates to a multi-gene panel for ex vivo molecular diagnosis of gastrointestinal stromal tumors and a kit comprising such a panel.

The invention relates to the technical field of assisted reproduction polygene detection, and discloses an oocyte occurrence and maturation disorder detection panel, a detection kit and application thereof, and the detection panel comprises mutation genes related to the oocyte occurrence and maturation disorder for detection, copy number variation genes and rearrangement event and deletion genes. According to the application, by detecting high-risk genes and mutation sites, the risk of occurrence and maturation disorder of the oocytes is predicted in combination with clinic, the development potential of the oocytes is evaluated, the in-vitro maturation strategy is optimized, and the clinical outcome of assisted reproduction is improved. The detection panel can be used for efficiently detecting gene mutation which has clinical diagnosis and treatment significance on oocyte occurrence and maturation disorders; the method has important guiding significance on genetic counseling, risk prediction, clinical diagnosis, precise treatment and potential personalized intervention measures of patients suffering from recurrent oocyte occurrence disorder, oocyte maturation disorder and low oocyte maturation rate and patients to be subjected to assisted reproduction technology treatment.

The invention provides a method for accurately and efficiently evaluating gene editing efficiency based on NGS data, a storage medium and electronic equipment, and belongs to the technical field of biological information processing. The method comprises the following steps: splitting NGS mixed pool sequencing data according to a bar code tag sequence, constructing a reference genome index and a dictionary, comparing NGS data of each sample with a reference genome, counting a comparison rate of a comparison result, constructing a comparison result index, converting the comparison result into a tsv format, and evaluating editing efficiency integration of a gene editing target site in each sample. The requirement for accurately and efficiently evaluating the gene editing efficiency based on NGS data is met. According to the method, the limitation of an existing tool on the size of a data file can be effectively avoided, the bar code label sequence can be customized for data splitting, and a high-accuracy comparison result is obtained by utilizing a mainstream comparison algorithm tool; and parallel operation is carried out by utilizing the advantages of the computing cluster, so that the aim of accurately and efficiently evaluating the gene editing efficiency of multiple genes and a large number of samples is fulfilled.

The invention provides a double-tandem base editor as well as a preparation method and application thereof, and belongs to the technical field of gene editing. The double tandem base editor disclosed by the invention is obtained by recombining a gRNA1-tRNA-gRNA2 unit component and an adenine base editor, the gRNA1 is designed for a target gene 1 and comprises a gRNA target specific spacer region and a gRNA skeleton of the target gene 1; the gRNA 2 is designed for a target gene 2 and comprises a gRNA target specific spacer region and a gRNA skeleton of the target gene 2. According to the double tandem base editor provided by the invention, different gene loci can be targeted only by changing 20bp gRNA, multi-gene editing on large animal cells is realized, accurate and safe multi-gene editing is realized under the condition of not introducing double-strand breakage, and the mutation efficiency is higher than that of a multi-plasmid co-transfection base editor.

The invention discloses a host gene combination for diagnosing acute bacterial and virus infection and application of the host gene combination. According to the present invention, with 100 related characteristic genes, the rapid diagnosis of acute bacterial and virus infection can be achieved; according to the detection result of the multi-gene combination, acute bacterial and virus infection, risks and types are diagnosed, clinical information and the like of patients are combined, more understanding is provided for clinical infection type cancer progression of the patients, accurate diagnosis and treatment are facilitated, and the clinical diagnosis and treatment efficiency is improved. Meanwhile, detailed data are provided for detection standardization of patients with acute bacterial and virus infection risks.

The present invention relates to transfer DNAs which can be used to produce a genetically modified plant that only has DNA from that species of plant. The present invention also relates to genetically modified plants with a transfer DNA of the invention which has been processed during integration and methods of making such plants. Also provided is a DNA construct and methods of making a multigene construct comprising five genes which confer upon a plant enhanced resistance to a wide variety of fungal pathogens.

The invention discloses a corynebacterium glutamicum engineering strain produced by L-glutamine as well as a construction method and application of the corynebacterium glutamicum engineering strain. The corynebacterium glutamicum engineering strain is prepared by integrating a glutamine synthase encoding gene glnA which is sourced from saccharomyces cerevisiae and has a tyrosine residue mutation at the 99th site on the surface and an sRNA-MicC-EcHfq regulation module of a targeted adenylyltransferase gene glnE into a vector plasmid; the method comprises the following steps: constructing a recombinant plasmid for removing adenosine acylation of glutamine synthase; and transferring the recombinant plasmid into corynebacterium glutamicum of which the odhA gene expression is controlled by a growth stage specific promoter Pcg2705, so as to obtain a corynebacterium glutamicum engineering strain for producing L-glutamine. According to the invention, through multi-gene synergistic modification and space-time metabolic flux optimization, the problem of'growth-synthesis' tradeoff in the prior art is solved, the fermentation efficiency is improved, the fermentation process is simple, and the application prospect is good.

The invention discloses a genetically engineered bacterium for high yield of L-alanine as well as a construction method and application of the genetically engineered bacterium, a dominant strain is obtained by knocking out an alanine racemase gene dadX and integrating coding sequences of alanine dehydrogenase from different sources, and the shake flask yield is 10.8 g / L; the activity of a corresponding mutant M18L / E75A / A78S / I266V is 4.3 times that of a wild type; 6 by-product related genes are knocked out, meanwhile, a multi-copy strain integrated with alanine dehydrogenase is constructed, and the yield of E.coli A-LB6 in a 5L fermentation tank is increased to 133.3 g / L. Through a dynamic regulation and control strategy, an oxygen-sensitive promoter Pomp * is adopted to replace a phosphoenolpyruvate carboxylase promoter, so that succinic acid byproducts are reduced by 85.8%, and the yield reaches 143.5 g / L after 48 hours of aerobic and anaerobic two-stage fermentation. And finally, by regulating and controlling the ATP synthetase gene cluster atp promoter, the yield of L-alanine in a 50 L fermentation tank reaches 151.3 g / L, and the saccharic acid conversion rate reaches 95.2%. The combination of polygene knockout and dynamic metabolism regulation effectively enhances the biosynthesis ability of alanine, and provides an efficient engineering strain for industrial production.

The embodiment of the invention provides a library construction method, a universal joint and a kit, relates to the technical field of synthetic biology, and is used for constructing a mixed library in which multiple gene segments are directionally connected. The library construction method comprises the following steps: providing a plurality of first initial gene segments; introducing cutting sequences to two ends of each first initial gene segment to obtain a plurality of second initial gene segments; introducing universal joints to two ends of each second initial gene segment to obtain a plurality of target gene segments; wherein the universal joint comprises an identification sequence; performing cutting and connection reaction on the plurality of target gene segments to obtain a mixed library; wherein the plurality of target gene segments are connected according to a set sequence. The library construction method is used for constructing the DNA mixed library.

Disclosed herein is a polynucleotide construct comprising one or more primary endonuclease recognition sequences upstream and downstream of a multiple gene editing site that comprises a plurality of secondary endonuclease recognition sequences. The primary endonuclease recognition sequences facilitate insertion of the multiple gene editing site into a host cell genome. The secondary endonuclease recognition sequences facilitate insertion of one or more exogenous donor genes into the host cell.

The invention discloses a construction method and application of a toxoplasma gondii polygene deletion vaccine strain. According to the invention, a TgBFD1 gene and a TgDAHPS gene are respectively knocked out from an ME49 [delta] 6pgdh1 insect strain through a gene editing technology, and a polygene deleted insect strain [delta] 6pgdh1 [delta] bfd1 [delta] dahps is successfully constructed; the strain can be subcultured in vitro, has the characteristics of no toxicity and no formation of tissue cysts in a host body, has better immune protection after immunization and inoculation, and can resist lethal infection of highly toxic strains. The toxoplasma gondii polygene deletion strain provided by the invention has better safety than ME49 delta 6pgdh1 strain, the toxicity of the toxoplasma gondii polygene deletion strain is obviously reduced, tissue cysts are not formed, the toxoplasma gondii polygene deletion strain can effectively resist infection of wild poisonous strains, the toxoplasma gondii polygene deletion strain has good immune protection capability, the defects that ME49 delta 6pgdh1 still has toxicity and can form tissue cysts are overcome, and the toxoplasma gondii polygene deletion strain has good application prospects. A method is provided for developing a safe and efficient toxoplasma gondii vaccine.

Disclosed herein a unique cell line system to generate a novel bovine adenovirus vector that provides more gene insertion capabilities and better immunogenicity for inserted antigens. The unique cell line is used for generating and growing of the new adenovirus vectors for gene delivery or recombinant vaccine production.

The present application relates to the field of gene editing technology, bioengineering and application of microbial fermentation, and discloses a Pichia pastoris high-efficiency gene editing system, a construction method thereof and application in a high-yield cordycepin engineering strain.The construction of the Pichia pastoris CRISPR-Cas9 gene editing system mainly includes the following steps: the histidine HIS4 nutritional deficiency back complementation is used to realize the resistance-free assembly of KhCas9; the sgRNA-tRNA array and the IIS type restriction endonuclease combination are used to simplify the gene editing operation and realize the simultaneous and high-efficiency editing of multiple genes; and the Brex27 domain is fused with KhCas9 to strengthen the homologous recombination efficiency. By using the gene editing system, through the optimization of methanol assimilation, 3'-AMP and adenosine precursor supply, ATP / NADPH energy supply and the like, the construction of a cordycepin engineering strain without resistance gene residues is realized for the first time, the gene editing efficiency of Pichia pastoris is greatly improved, and a powerful tool is provided for the industrialized production of cordycepin.

The invention relates to the technical field of gene vectors, and discloses a multi-target multi-gene stable silencing vector and an application thereof in treating diabetes mellitus, and a multi-gene multi-target gene modification combined element is safely and efficiently integrated at a gene safety parking point of a second intron region of a CCR5 gene in a genome in a reverse fixed point manner by using a non-viral vector system; therefore, the effect of continuously silencing expression of a plurality of target genes such as MSTN, GRB10 and PTBP1 for a long time is achieved, and aging of mesenchymal stem cells cultured in vitro is remarkably inhibited. When the mesenchymal stem cells modified by the multi-target multi-gene stable silencing vector are infused, the activity is high, the chronic inflammatory microenvironment is effectively inhibited, and the senescence process is delayed; moreover, proliferation and differentiation of pancreatic beta cells are promoted, insulin secretion is enhanced, insulin resistance is effectively resisted, and diseases such as diabetes mellitus are prevented.

The invention discloses a method for cultivating a heat-resistant broad-spectrum durable rice blast-resistant rice breeding material based on polygene polymerization. According to the method, a high-heat-resistance and high-dominance indica-japonica hybrid indica rice restorer line is selected as a recurrent parent, a breeding intermediate material for polymerizing complementary broad-spectrum rice blast resistance genes Pigm and Pi-jx is used as a donor parent, and polymerization of the heat-resistance and broad-spectrum lasting rice blast resistance genes Pigm and Pi-jx is realized through a method of combining molecular breeding with conventional breeding. Meanwhile, by selecting representative strains to carry out artificial inoculation identification of seedling blast and panicle blast, natural induction identification of a rice blast retransmission area and heat resistance identification, and by combining main agronomic trait evaluation in the whole growth period, a heat-resistant broad-spectrum durable rice blast-resistant breeding material is obtained. The breeding material bred by using the method disclosed by the invention has relatively strong heat resistance and broad-spectrum and lasting rice blast resistance, and the breeding process of heat-resistant and rice blast-resistant rice varieties is accelerated.

This invention belongs to the field of biomedical technology, specifically relating to the application of DNA damage response gene-related SNPs in the treatment and prognosis of acute myeloid leukemia (AML). This invention is the first to discover that single nucleotide polymorphisms (SNPs) in the DNA damage response pathway (focusing on three major functional modules: damage sensing, signal transduction, and DNA repair) are closely related to the clinical characteristics, treatment response, and survival prognosis of AML. This lays the foundation for constructing a multi-gene risk model based on the DNA damage response pathway and provides theoretical support for integrating germline SNP maps into precision medicine strategies for AML.

The invention discloses a mouse multi-gene collaborative insertion and knockout system based on multi-data coupling, and relates to the technical field of bioengineering, and the mouse multi-gene collaborative insertion and knockout system comprises a multi-gene collaborative targeting vector, a high-fidelity Cas9 protein expression unit, a homologous recombination template library, a multi-modal efficiency prediction module, a dual-fluorescence Cre report unit and an off-target inhibition element. According to the method, vector silencing is avoided by connecting the nuclear matrix attachment region in series with the gRNA expression cassette, and the multi-gene collaborative editing efficiency is improved to gt by combining homologous arm optimization design and a machine learning driven efficiency prediction model; 80%; cas9 fusion deaminase and a glycosylase inhibitor are combined with miRNA-mediated off-target inhibition, so that off-target sites of a whole genome are reduced to be less than or equal to 1 / sample; real-time monitoring of tissue specific expression is achieved by means of a double-fluorescence Cre reporting system, space-time regulation and control application is supported, and a high-reliability tool is provided for complex gene interaction research.

Provided are a prime editing system-based method for achieving inversion editing of a large genomic fragment and a composition used. By means of the efficient multi-gene prime editing protein construct ePPEplus in plants, the inversion of large fragments in plants is achieved by means of designing paired pegRNAs (pegL and pegR), and then by means of processes such as reverse transcription, DNA double-stranded annealing, DNA strand synthesis, and repair. The pegL comprises RTT-L, and the pegR comprises RTT-R. The nucleotide sequence of RTT-L is identical to that of strand A at the right terminus of a double-stranded DNA to be inverted. The nucleotide sequence of RTT-R is identical to that of strand B at the left terminus of said DNA. By means of testing in the monocotyledonous plant wheat and dicotyledonous plants Nicotiana benthamiana and tomato, it is indicated that the method is an effective method for mediating precise inversion of large-fragment DNA in plants.

The invention discloses a rice OsYUC12 / 13 / 14 polygene mutant as well as an encoding protein and application thereof. By simultaneously destroying biological functions of the encoding protein of three genes OsYUC12 / 13 / 14, the length of rice grains can be remarkably increased, and the content of rice apparent amylose can be reduced; the gene OsYUC12, the gene OsYUC13 and the gene OsYUC14 are knocked out at a fixed point at the same time through the CRISPR / Cas9 technology, a function deletion mutant is obtained, the length of rice grains is remarkably increased, the appearance quality of rice is improved, meanwhile, the apparent amylose content is remarkably reduced, and the eating quality of the rice can be improved; the new function of the rice YUCCA gene family OsYUC12 / 13 / 14 mutant provided by the invention provides a candidate gene for plant breeding, and also provides a potential application value in the aspect of improving the rice yield and quality.

The present invention discloses a method for detecting tumor multi-genes based on hyperspectral images, which belongs to the technical field of mutation sample detection, and specifically includes: obtaining tumor tissue sections of different carcinogenic driver gene mutation states after H&E staining; photographing tumor tissue sections by a hyperspectral camera, obtaining hyperspectral images of tumor samples and preprocessing; building a global feature extractor based on S3Anet; building a tumor multi-gene detection model and using multi-task learning training; collecting tumor samples to be detected, and obtaining hyperspectral data of the samples to be detected by a hyperspectral camera; inputting the hyperspectral data into a trained S3ANet model for feature extraction, and then inputting it into a trained multi-gene detection model to obtain the gene detection results of the tumor samples. The beneficial effects of the present invention are: the present invention effectively reduces the dependence on traditional pathological diagnosis experience and gene sequencing, significantly improves the accuracy, reliability and generalization ability of detection, and reduces the detection cost.

The application belongs to the technical field of molecular biology and medicine, and discloses a methylation marker for detecting the benignity and malignancy of thyroid nodules and an application. 780 new methylation sites are identified, 273 tumor occurrence and development related genes are associated, and the Fc gamma R-mediated phagocytosis signal pathway is involved. The methylation marker for detecting the benignity and malignancy of thyroid nodules comprises multiple genes and multiple methylation sites. The methylation level of multiple genes and multiple sites is detected, the problem of low and unstable methylation signal of single gene or single site is overcome, and thus the sensitivity and specificity of detection are improved.

The Quadruplet Expanded DNA (QED) eukaryote genetic code comprising twenty nondegenerate QED codons encode proteins (the protein-encoding codons), and thirty-five nondegenerate QED codons (the noncoding codons) being highly correlated with cis-regulatory elements control and regulate transcription, alternate splicing, and polymerization in eukaryotic protein synthesis using canonical amino acids. The QED eukaryote genetic code is an advancement to gene therapeutics that allows for the correction of dysfunctional proteins. Additionally, the QED eukaryote genetic code is further applicable for changing paradigms relating to identifying cures for monogenic rare, multigene cancer, and neurodegenerative diseases.