26 results about "Essential gene" patented technology

The invention belongs to the technical field of essential gene prediction, and particularly relates to an essential gene prediction method based on DNA large model and time-frequency domain deep learning fusion, and the method comprises the steps: taking a domain DNA large model as a core representation layer, and obtaining special gene representation through cross-species corpus pre-training and task fine tuning; a T-Block and F-Block dual-channel time-frequency fusion structure is adopted, and the local dependence and long-range regulation relation of a gene sequence is synchronously captured by expanding DFT (Discrete Fourier Transform), complex value attention and iDFT (Initial Discrete Fourier Transform) conversion; designing an efficient modeling reasoning scheme of sliding window slices and gene-level aggregation aiming at an ultra-long sequence; in combination with class imbalance and a noise robust training strategy, cross-cell line / cross-platform transferable threshold output is realized through temperature scaling calibration, an uncertainty quantization and structured interface is matched, and drug target screening and experimental design decision are supported. The system supports the realization of multiple programming languages, and can complete low-delay end-to-end reasoning in a conventional hardware environment.

The invention discloses a screening and expression platform construction method for non-essential gene loci of corynebacterium glutamicum, and belongs to the technical field of biology. The center sites CGLRS02940, CGLRS03085, CGLRS08555, CGLRS09020 and CGLRS13560 of the corynebacterium glutamicum identified by the invention have the remarkable advantages that when gene knockout or knockin operation is carried out, the sites do not interfere with the normal physiological metabolism process of strains, and stable expression of exogenous genes can be guaranteed; meanwhile, the gene editing efficiency is relatively high, and an editing target with high universality and high reliability is provided for genetic engineering modification of corynebacterium glutamicum.

The invention belongs to the technical field of directed evolution modification of protein, and particularly relates to an engineered bacillus subtillis chassis cell for inserting unnatural amino acid, which is formed by uniformly replacing termination codons UAG or UGA of 57 essential genes of bacillus subtillis with UAA. On the basis, two sets of mutually orthogonal translation systems are introduced to decode the UAG and the UGA respectively, and finally site-specific insertion of the two non-natural amino acids at the preset site of the target protein is realized; a stable and reliable cell platform is provided for obtaining protein molecules carrying two different chemical functional groups and carrying out multi-site refined modification research.

The invention provides a genetic engineering strain, a construction method of the genetic engineering strain and application of the genetic engineering strain in biosynthesis of 5-hydroxytryptophan. According to the engineering bacterium, four key modules required by 5-HTP synthesis are sequentially introduced into a host genome in a genome integration mode through a CRISPR technology, and the four key modules comprise an aroGfbr-ppsA-tktA module for enhancing chorismate precursor supply, a trpEfbr-trpG module for catalyzing o-aminobenzoic acid generation, a salABCD gene cluster responsible for hydroxylating o-aminobenzoic acid into 5-hydroxy o-aminobenzoic acid, and a salABCD gene cluster responsible for hydroxylating 5-hydroxy o-aminobenzoic acid into 5-hydroxy o-aminobenzoic acid. And a trpDCBA module for converting the intermediate into the final product 5-HTP. The modules are respectively integrated into a plurality of non-essential gene regions, so that stable genetic expression is realized, and a complete metabolic pathway can be operated without plasmids or inducers. The strain can continuously accumulate 5-hydroxytryptophan (5-HTP) under a glucose fermentation condition, and has the advantages of simplicity and convenience in construction, high hereditary stability, low production cost, suitability for industrial-scale fermentation and the like.

The application discloses a tumor cell line drug response prediction method integrating multiple omics and essential gene information, relates to the field of tumor drug response prediction, and is a DROEG prediction method (Drug Response based on Omics and Essential Genes). Four kinds of omics data, including gene expression data, copy number variation data, methylation data and somatic mutation data, and CRISPR gene effect data (CERES Score) are introduced to construct a drug response model, and the effect of the model is comprehensively evaluated by a qualitative evaluation method and a quantitative evaluation method. The application introduces CRISPR essential gene information, establishes a drug response prediction model, is more accurate than existing methods, has the functions of quantitative and qualitative prediction and evaluation, is more suitable, and is more flexible to operate.

The present invention relates to a fusion polypeptide comprising an IL10 polypeptide, which can be prokaryotically expressed and secreted in bacterial cells. The present invention also relates to a modified bacterium having anti-tumor activity in tumors and the ability to prokaryotically express and secrete an IL10 polypeptide, wherein the bacterium comprises an essential gene expression cassette controlled by an obligate hypoxia-inducible promoter, and the bacterium is deficient in at least one gene, or its functional expression product, involved in or regulating an endogenous antioxidant stress response pathway. The present invention also relates to a pharmaceutical composition comprising the modified bacterium and its anti-tumor use.

The present invention provides a method for producing a target protein by culturing cells, the method being capable of stably maintaining a target plasmid without using an antibiotic. The present invention relates to a method for producing a target substance, the method including a step for culturing cells that are transformed with a target plasmid (A), wherein: each of the transformed cells includes the target plasmid (A) and a temperature-sensitive plasmid (B) and are obtained by a method that includes a step for culturing cells in each of which the function of an essential gene on the chromosome has been reduced at a temperature at which the temperature-sensitive plasmid (B) is eliminated; the target plasmid (A) includes the essential gene and a target substance gene and does not include an antibiotic-resistant gene; and the temperature-sensitive plasmid (B) includes the essential gene and the antibiotic-resistant gene.

The invention relates to the technical field of plant genetic engineering, in particular to a gene down-regulation method based on regulation of precursor mRNA (messenger Ribonucleic Acid) splicing efficiency, which comprises the following steps: firstly, identifying at least one intron 5'terminal splicing site containing a GT sequence in a target gene; aiming at the site, designing a guide RNA (Ribonucleic Acid) sequence capable of targeting the downstream fourth or fifth nucleotide, and verifying the cleavage activity of the guide RNA sequence; constructing a gene editing vector by using the guide RNA sequence which is verified to be effective and a Cas9 nuclease coding gene; introducing the vector into a plant cell, carrying out genetic transformation and regeneration to obtain a transformed plant, and screening out a mutant which is subjected to site-directed editing at a target site through DNA (Deoxyribose Nucleic Acid) sequencing; and finally, extracting RNA (Ribonucleic Acid) of the mutant, reversely transcribing into cDNA (Complementary Deoxyribonucleic Acid), and analyzing transcript change through PCR (Polymerase Chain Reaction) amplification and gel electrophoresis. According to the invention, stable, heritable and accurate knock-down of gene expression is realized, and a new tool is provided for essential gene function research and crop character improvement.

The invention provides a big language model and multi-modal data-based complex character gene mining method and system, and belongs to the technical field of gene data analysis, the method comprises the following steps: obtaining a statistical data set containing multi-modal data of different bean populations; annotating all genes in the statistical data set based on a reference genome to obtain a whole genome semantic map; if the large language model receives a site of a target significance character, calculating a comprehensive score of each gene at the site by using the large language model based on the whole genome semantic map and the site, and determining a complex character gene based on the comprehensive score of each gene; and if the large language model does not receive the locus of the target saliency character, determining candidate genes by using the large language model based on the target saliency character, and determining complex character genes from the candidate genes by using the large language model based on the whole genome semantic map. According to the invention, the most essential gene influencing the target significance character can be positioned.

The invention relates to the technical field of biology, in particular to application of a bombyx mori sorbitol dehydrogenase gene BmSdh2 in relieving diapause of bombyx mori. The nucleotide sequence of the silkworm sorbitol dehydrogenase gene is as shown in SEQ ID NO: 1 or SEQ ID NO: 2. The gene editing technology is utilized to realize successful knockout of the bombyx mori diapause essential gene, and mutation of a corresponding target spot on a genome is detected. PCR amplification and genome sequencing are used for detection of gene mutation, the obtained homozygous mutant material offspring does not enter diapause, the diapause-controllable practical silkworm strain is constructed, and large-scale factory breeding can be realized.

The invention belongs to the technical field of biological genetic engineering, and particularly relates to a corn kernel development regulation gene ZmPPR299, an encoding protein thereof, an SNP (Single Nucleotide Polymorphism) site, a molecular marker and application of the molecular marker, the gene ZmPPR299 is located in a corn chromosome 5, and functional verification shows that the gene encodes a PPR protein located in chloroplast, and the molecular marker can be used for regulating the development of corn kernels. The chloroplast gene is an essential gene for regulating corn kernel development and chloroplast functions; the mutation of the gene can cause the lagging of embryo and endosperm development, the reduction of basal transfer layer cell proliferation, and the insufficient filling of starch grains and protein bodies, and finally, the results show that the grains shrink, the hundred-grain weight is obviously reduced, and the leaves of seedlings lose green. The invention also develops a specific CAPS molecular marker and a detection primer thereof based on a single base mutation site of the gene, and the genotype of the mutant can be rapidly and accurately identified. The gene resource and the molecular marker provided by the invention have important application values in analysis of a corn kernel development regulation network, development of molecular assisted breeding and creation of high-yield and high-quality new corn germplasm.

The invention relates to the technical field of biological medicines, in particular to application of a DBC1 gene in preventing or treating alopecia diseases, and any one of the following applications of the DBC1 gene is as follows: preparing a medicine or a health care product for maintaining the number or function homeostasis of Treg cells; preparing medicines or health-care products for maintaining or increasing the number of the hair follicle stem cells; preparing medicines or health-care products which support normal growth of hair follicles and are complete in structure; preparing medicines or health-care products for driving hair regeneration; and preparing medicines or health-care products for maintaining the stability of the peripheral immune microenvironment of the hair follicle stem cells. The invention discloses a key effect of the DBC1 gene in hair regeneration, and the DBC1 is confirmed to be a necessary gene for promoting hair regeneration through a gene knockout animal model, systemic knockout and specific knockout for the first time.

The invention provides a cDNA (complementary deoxyribonucleic acid) clone plasmid of an avian infectious bronchitis virus (IBV) for expressing secretory luciferase as well as a virus strain and a vaccine obtained by rescuing, and belongs to the technical field of biological medicines. The invention provides a construction method of cDNA (complementary deoxyribonucleic acid) clone plasmids of IBV (infectious bursal virus), which is completed by a one-step method based on yeast homologous recombination in a segmented amplification mode. The IBV infectious cDNA clone constructed by the method has good stability, an IBV genome can be transformed on a gene level by directly utilizing in-vitro homologous recombination, and the efficiency of constructing a recombinant virus is improved. According to the invention, a non-essential gene of the infectious cDNA clone is replaced by a Gluc reporter gene, and the obtained infectious cDNA clone directly transfects mammalian cells to rescue recombinant viruses. The recombinant virus is constructed through a virus rescue method, the growth characteristic of the recombinant virus is basically consistent with that of a parent virus, and genetic stability is kept.

The application discloses a data processing device and method for predicting essential genes of non-model organisms in the field of bioinformatics and application. The technical problem to be solved by the application is how to screen essential genes of non-model organisms. In the application, the protein sequences of a target species and the protein sequences of n model organism species are respectively based on annotation to obtain the corresponding labels of each protein; then the protein sequences and the labels are embedded into a protein language model to output embedding vectors; based on the embedding vectors, a residue contact map is constructed for each protein; the features of each amino acid residue in the residue contact map are modeled and integrated using a graph neural network to output a whole protein embedding vector; all protein embedding vectors are input into a classifier model for training and using an activation function to output essential probability values of protein genes; and based on the classifier model, essential probability values P1 of the genes of the proteins are obtained. The application can be applied to agricultural disease prevention and control and biological pesticide development.

An improved baculovirus expression system for enhanced heterologous protein expression in hosts selected from Bombyx mori larvae, pupae, primary cells, or established insect cell lines is disclosed. The expression system is derived from a wild-type strain of Bombyx mori nucleopolyhedrovirus (BmNPV) by deletion or inactivation of non-essential genes, of the BmNPV genome. The system includes novel transfer vectors designed for targeted gene modification and incorporation of a bacterial origin of replication and an insertion site for a gene of interest (GOI). This engineered bacmid maintains infectivity while offering enhanced genetic stability and significantly improved heterologous protein expression in insect cells, larvae, or pupae. Methods for constructing this bacmid via homologous recombination and using it for high-level protein production are also described.

The invention belongs to the technical field of gene editing, and particularly discloses a zymomonas mobilis genome editing method and application. The method comprises the following steps: by taking zymomonas mobilis as a chassis, carrying out fusion expression on programmable nuclease HNH and Cas8 (ZMO0682), and constructing a CRISPR-Cas8-HNH system; further, the cas1 and cas2 / 3 in the strain are knocked out through homologous recombination, and a CRISPR-Cas8-HNH [delta] Cas1 [delta] Cas2 / 3 system is obtained; the system solves the problems that a Zymomonas mobilis exogenous CRISPR-Cas system is high in cytotoxicity, no transformant exists or few transformants exist when essential genes are knocked out by an endogenous system and an exogenous system, and the essential gene knockout efficiency is low. The engineering strain has the advantages of enhancing the knockout efficiency of essential genes, greatly increasing the number of transformants when the essential genes are knocked out, greatly increasing the editing efficiency and the like.

The invention relates to a method of altering a genome of a microorganism with a non-replicating construct and a regulatable inhibitor of RNA or protein synthesis that targets an essential gene of the microorganism. The invention further relates to a non-replicating targeting construct comprising a regulatable inhibitor of RNA or protein synthesis that targets an essential gene of the microorganism.

The invention relates to a grass carp cell membrane protein gene CRISPR (clustered regularly interspaced short palindromic repeats) activated plasmid library. The library comprises CRISPR activated plasmids for targeted activation of 3498 candidate grass carp cell membrane protein genes; the invention further relates to a recombinant lentivirus capable of infecting fish cells and a packaging system of the recombinant lentivirus. The invention provides a CRISPR (clustered regularly interspaced short palindromic repeats) activation library aiming at related genes of grass carp cell membrane protein for the first time, and further provides recombinant lentivirus capable of infecting fish cells and a packaging system of the recombinant lentivirus. Protein on a grass carp cell membrane is identified through experiments, genes which exist in genome prediction but are not actually expressed can be excluded, and a library which accurately reflects related genes of grass carp cell membrane protein is formed. The CRISPR activation library provided by the invention avoids the situation of cell death after essential genes in a gene knockout library are knocked out, can comprehensively aim at essential or non-essential genes for cell survival, and can be widely applied to scenes such as virus receptor identification and / or gene function research and / or drug screening.

An improved baculovirus expression system for enhanced heterologous protein expression in hosts selected from Bombyx mori larvae, pupae, primary cells, or established insect cell lines is disclosed. The expression system is derived from a wild-type strain of Bombyx mori nucleopolyhedrovirus (BmNPV) by deletion or inactivation of non-essential genes, of the BmNPV genome. The system includes novel transfer vectors designed for targeted gene modification and incorporation of a bacterial origin of replication and an insertion site for a gene of interest (GOI). This engineered bacmid maintains infectivity while offering enhanced genetic stability and significantly improved heterologous protein expression in insect cells, larvae, or pupae. Methods for constructing this bacmid via homologous recombination and using it for high-level protein production are also described.

The invention discloses application of a magnaporthe oryzae squalene cyclooxygenase gene MoERG1 in prevention and treatment of magnaporthe oryzae, and belongs to the technical field of agricultural biology. The invention proves that the squalene cyclooxygenase gene MoERG1 of the magnaporthe oryzae is a key essential gene for environmental adaptability and pathogenicity of the magnaporthe oryzae. The gene is specifically silenced by an RNA interference technology, so that the completeness of cell walls of pathogenic bacteria can be effectively damaged, the tolerance of the pathogenic bacteria to oxidative stress and hypertonic stress is remarkably weakened, the yield of conidia and the formation rate of a key infection structure-appressorium of the pathogenic bacteria are greatly reduced, and finally, the pathogenic ability of the pathogenic bacteria to rice is completely lost. The core function of the squalene ring oxygenase gene MoERG1 as a multi-effect regulation target is disclosed from the molecular level. In order to solve the industrial problems that the existing single-target bactericide is easy to generate drug resistance and the traditional biocontrol effect is unstable, the invention provides a brand-new effective solution with a more fundamental action mechanism.

The invention relates to the technical field of biology, in particular to an escherichia coli gene composition clustering design and synthesis method. According to the technical scheme, the method mainly comprises the following steps: performing de novo design synthesis on clustering in a whole genome range of escherichia coli, and dividing the escherichia coli into an essential gene cluster, a non-essential gene cluster A and a non-essential gene cluster B; in a design and assembly process, a loxPsym site is inserted into a non-essential gene cluster A, and a vox site is inserted into a non-essential gene cluster B, so that the flexibility of a chassis is improved, structural variation in a whole genome range can be quickly generated, the coverage range is wider, and the gene operation scale is larger.

More particularly, the present invention relates to a modified bacterium having anti-tumor activity, the bacterium comprising a hypoxia regulatory lipopolysaccharide (LPS) gene expression cassette comprising a gene involved in LPS biosynthesis or transport or a gene encoding a mural lipoprotein controlled by a first strict hypoxia inducible promoter. The bacterium further comprises an essential gene expression cassette controlled by a strict hypoxia inducible promoter, and the bacterium lacks at least one gene participating in or regulating an endogenous anti-oxidative stress response pathway or a functional expression product thereof. The invention also relates to a pharmaceutical composition containing the modified bacterium and an anti-tumor application thereof.

This invention discloses a virus that specifically kills tumor cells and a tumor therapeutic drug, relating to the field of biotechnology. The virus is a recombinant oncolytic virus with a foreign promoter located upstream of the virus's essential gene, replacing the natural promoter of the essential gene to drive its expression in tumor cells while remaining unexpressed in normal cells. This virus can specifically kill various tumor cells, exhibiting high killing efficiency against tumor cells while remaining safe for non-tumor cells.

The present application relates to a kind of orthogonal type DNA self-replicating vector and its use in mammal to express gene of interest.The vector includes the expression cassette of the replication essential gene of type I herpes simplex virus and replication initiation sequence, wherein the replication essential gene of type I herpes simplex virus is UL9, UL29, UL30, UL42, UL5, UL8 and UL52, and the replication initiation sequence is OriS and / or OriL.

The invention discloses a method for improving production of citrulline by Escherichia coli, which comprises the following steps: knocking out one, two or three of the following three unnecessary gene sequences in an Escherichia coli genome for producing L-citrulline: a sequence DLP12 located between genome coordinates 564755-586065, a sequence Part4 located between genome coordinates 3110645-3134692, a sequence DLP12 located between genome coordinates 564755-586065 and a sequence Part4 located between genome coordinates 3110645-3134692; and the sequence CP4-57 is located between the genome coordinate 2755941 and the genome coordinate 2777971. The fermentation yield of the L-citrulline can be increased by 27.42% by knocking out the three gene sequences at the same time, and the method has industrial application prospects.