20 results about "Eukaryote" patented technology

The invention discloses an influenza C virus HEF protein polyclonal antibody preparation method, which comprises: carrying out signal peptide prediction, antigen epitope screening and hydrophobicity analysis through bioinformatics, constructing a recombinant prokaryotic expression vector pET32a-HEF by using a seamless cloning technology, and efficiently expressing the HEF protein in Escherichia coli to obtain the influenza C virus HEF protein polyclonal antibody. The inclusion body protein yield is improved by optimizing conditions; and purifying by nickel column affinity chromatography to obtain high-purity recombinant protein, emulsifying the high-purity recombinant protein and a Freund's adjuvant, immunizing a Japanese white rabbit, immunizing for three times, and collecting high-titer serum. The method breaks through detection limitation caused by antigen tag concealment in a eukaryotic expression system, the obtained polyclonal antibody is high in specificity, the eukaryotic expression HEF protein can be accurately recognized, and a key tool is provided for ICV virus-like particle identification, vaccine research and development and diagnosis technologies. Compared with a traditional method, the scheme is simple and convenient to operate, low in cost and high in antibody titer, and has remarkable application value.

A method of selectively binding DNA of bacterial origin comprising the step of contacting a sample solution containing DNA of bacterial origin with a protein comprising the TgMcrBΔ185N-terminal domain of Thermococcus gammatolerans McrB protein or a derivative thereof which has been immobilised into a substrate whenever the solution further contains DNA of eukaryotic origin at a concentration of more than 1-100 times that of the DNA of bacterial origin.

The present invention relates to adenine base editors (ABEs) and components thereof. The present invention also relates to complexes comprising an adenine base editor (ABE) and a guide RNA in a functionally associated form. The present invention further relates to a nucleic acid molecule encoding an ABE / guide RNA, an expression construct or vector comprising a nucleic acid sequence encoding an adenine base editor and / or a nucleic acid sequence encoding a guide RNA. The present invention further relates to a cell comprising an adenine base editor (ABE) and a method for adenine base editing of a target site in a genome of interest in at least one cell of a prokaryotic or eukaryotic organism, including bacterial and archaeal organisms. In addition, the present invention relates to various methods, kits and uses related to the provided ABEs.

The invention relates to application of DNA polymerase in long-fragment precise gene editing and construction in a plurality of human-derived cell lines. The invention belongs to the technical field of biological medicine. A novel gene editing system MMEJ-Pol theta is created by applying DNA polymerase Pol theta and a key functional structure domain, the accuracy, safety and editing efficiency of long-fragment gene editing in a eukaryotic genome are remarkably improved through a micro homologous end linking method, and an effective tool is provided for transformation application of the gene editing system in gene therapy and cell therapy.

The present invention provides a method and apparatus for identifying horizontally transferred sequences from bacterial genomes in recipient insect genomes, relating to the field of biotechnology. The method provided by the present invention for identifying horizontally transferred sequences from bacterial genomes in recipient insect genomes more comprehensively and accurately identifies and analyzes fragments horizontally transferred from a certain type of bacteria to the genome of this species at the whole genome level; based on the horizontally transferred sequence fragments, horizontally transferred genes, gene fragments, and non-coding region fragments are further identified. This provides a comprehensive analysis of horizontal gene transfer between the genomes of a certain eukaryote and a certain type of bacteria, laying a solid, comprehensive, and reliable foundation for further research into the effects of horizontally transferred genes (fragments) on eukaryotes.

The present invention relates to the technical fields of bioinformatics and molecular cytogenetics, and particularly relates to a method and application for chromosome centromere counting based on Nuf2 antibody. The present invention comprises the following steps: performing chromosome centromere counting on the Nuf2 antibody and the centromere-specific protein Nuf2 through an immunofluorescence reaction, so as to directly observe the number of chromosome centromeres, and thus efficiently and rapidly determine the number of chromosomes. In addition, in view of the high conservation of the centromere-specific protein Nuf2 in eukaryotes, the present invention also provides the application of this method in identifying the ploidy of plants. The operation is simple, the cost is low, the efficiency of ploidy identification is significantly improved, and a general technical platform is provided for genetic breeding.

Rhizobial infection is the key process for initiating symbiotic nitrogen-fixing root nodules in legumes, which requires specific recognition of signal molecules produced by the bacteria and their hosts. Here, it is established that rhizobial tRNA-derived small RNA fragments (tRFs) are crucial signal molecules modulating host nodulation, which uncovers a bacterial small RNA-mediated mechanism for prokaryote-eukaryote interaction. Transgenic plants are also provided that express a construct encoding rhizobial-derived tRNA, which is subsequently cleaved to produce artificial tRFs. Constructs and methods of producing the same are also provided, as well as modifications for repressing a mechanism for the negative regulation of nodulation present within plants.

An approach for differentially isolating eukaryotic (plant and animal) DNA from bacterial DNA prior to sequencing using a combination of size exclusion-based separation and differential cell lysis. The method of the present invention exploits the differences of the cellular size and components of each type of organism to be separated. The composition and nature of the cell wall of plant cells, enzymatic sensitivity of bacterial and animal cells and overall size difference of bacterial and plant / animal cells allows one portion of a mixed sample to be lysed while retaining the integrity of the remaining organisms. Separation of one phylogenetic component then permits the remaining components to be extracted with minimal contribution from the preceding component. The separation of DNAs from differing contributing kingdoms in an unknown sample increases interpretability through decreasing complexity in subsequent sequencing applications.

The application discloses a pig IZUMO2 eukaryotic protein and a preparation method and application of a monoclonal antibody of the pig IZUMO2 eukaryotic protein. The full-length sequence of pig IZUMO2 nucleotide CDS is verified, a research method for removing the transmembrane structure of the pig IZUMO2 nucleotide CDS and only expressing the extramembrane sequence of the pig IZUMO2 nucleotide CDS in eukaryotes is designed, and the IZUMO2 protein is expressed and secreted by CHO-K1 cells. The protein expressed by the method is close to natural protein, has biological activity and is simple to operate. The qPCR and ELISA tests verify that the IZUMO2 protein can reduce the expression of inflammatory factors in RAW264.7 cells induced by LPS, and has a certain anti-inflammatory capacity. It is found through experiments that the IZUMO2 protein can promote the capacitation of boar sperm, and it is proved that the protein can be applied to the preparation of a reagent for promoting the capacitation of boar sperm.

The invention belongs to the technical field of forensic medicine and biological medicine, discloses a method for forensic medicine identification of corpses in water based on metagenomics markers and application, and aims to solve the problems that existing drowning diagnosis is insufficient in accuracy and the deduction of post-death submerging time (PMSI) is limited. The method comprises the following steps: collecting a corpse lung tissue in water and a water sample in a corresponding water area, and obtaining a microbial DNA sequence by adopting a metagenome sequencing technology; the method comprises the following steps: processing data through bioinformatics analysis (such as fastp quality control, host sequence removal by KneadData and Kraken2 species annotation), and selecting and screening species level microbial markers in combination with a random forest algorithm and Boruta characteristics; and respectively establishing a drowning diagnosis model and a PMSI inference model based on the screened markers, and verifying the performance of the models through indexes such as AUC and MAE. Complete microbial communities of bacteria, eukaryotes (including fungi), archaea and viruses in lung tissues of corpses in water are comprehensively analyzed for the first time, 17 bacterial markers and 9 eukaryote markers are screened out for drowning diagnosis (verification experiment bacterial model AUC = 1, the accuracy rate is 89.29%, eukaryote model AUC = 0.95, the accuracy rate is 87.5%), and 17 markers are screened out for PMSI inference (integrated model MAE = 0.66 days). According to the method, the species resolution reaches the species level, the method is not influenced by amplification bias, the method is suitable for different water area environments, and an efficient and objective forensic medicine tool is provided for identifying the cause of the dead body in water and deducing the submerged time after the dead body.

A cross-kingdom platform for the delivery of therapeutic agents to target cells. The system maintains the transport and uptake functions of the Inv while altering targeting from β1 integrin to other proteins expressed on the surface of target eukaryotic organisms (i.e., cell surface proteins) or chemical moieties expressed on the surface of target eukaryotic cells (i.e., cell surface chemical moieties) by genetically replacing D4 and D5 of the Inv with binding domains from heterologous proteins. These heterologous proteins can be derived from bacterial, fungal, animal, or viral genomes. This engineering results in the construction of chimeric Inv proteins in which D1-D3 (i.e., non-binding domains) are fused in frame to alternative binding domains from heterologous proteins. The alternative binding domains interact with different cell surface proteins or chemical moieties, which can sometimes be referred to as receptors, on the surface of eukaryotic cells, thereby allowing specific targeting of the Inv to cells independent of endogenous β1 integrin binding.

Synthetic genetic platforms in eukaryote cells (such as yeast) is described. A representative synthetic genetic platform includes a eukaryote cell genetically modified to express an auxin receptor, an auxin response factor, and a reporter, as well as a fusion construct including a Lis1 Homology (LisH) domain fused to an auxin-responsive protein. The synthetic genetic platforms can be used, for instance, to understand developmental and pathological LisH domain variants, and to test bioactive molecules for LisH domain activity.

The invention discloses preparation and application of oligopeptide immune carrier protein, and belongs to the technical field of biology. The method comprises the following steps: fusing a gene sequence for coding oligopeptide and a gene sequence for coding ferritin through a chemical method by utilizing the characteristics that oligopeptide molecules are small in mass and ferritin is self-assembled to form a polymer, and then cloning the fused gene into a prokaryotic biological carrier for expression; the recombinant protein is purified by using an expression system which is not limited to prokaryotes such as escherichia coli and eukaryotes. The protein has the advantages of low preparation cost, strong immunogenicity and the like, and lays a foundation for obtaining a high-affinity anti-oligopeptide nano antibody.

The invention discloses a T cell receptor for specifically recognizing mycobacterium tuberculosis antigen peptide as well as a preparation method and application of the T cell receptor. According to the invention, HLA-A * 0201 positive pulmonary tuberculosis patient PBMC is respectively stimulated by tuberculosis antigens CFP10 and Ag85B, CD8 + T cells are subjected to flow sorting, TCR alpha / beta chains are paired, corresponding HLA-A * 0201 restrictive TCR protein is successfully obtained, and eukaryotic expression and purification are carried out. The TCR protein can significantly inhibit proliferation of mycobacteria in THP-1 macrophages, shows that CFU is reduced and high content fluorescence intensity is weakened, and significantly improves expression of inflammatory factors such as IL-1beta, TNF-alpha, IL-6, IL-12 and the like. The tuberculosis antigen-specific TCR protein is proved to effectively inhibit intracellular mycobacterium tuberculosis replication by regulating the polarization state of macrophages and an inflammatory signal network for the first time, and a new strategy is provided for TCR-based tuberculosis immunotherapy.

The invention discloses Yarrowia guilliermondii, a preparation method of an inactivated vaccine of the Yarrowia guilliermondii and application of the Yarrowia guilliermondii in prevention and treatment of alcoholic liver diseases, and belongs to the field of microorganism eukaryotes, the preservation date of the Yarrowia guilliermondii is August 18, 2025, and the preservation number is CGMCC NO. According to the present invention, the vaccine is prepared by separating and inactivating the Yarrowia guilliermondii ML001, and the vaccine can significantly reduce the alcohol-induced ALT and AST levels in mice, reduce the liver lipid accumulation, reduce the fatty acid synthesis genes Fasn and Acly, and increase the fatty acid oxidation genes Ppara and Cpt1a so as to effectively inhibit the lipid synthesis and activate the fatty acid oxidation, moreover, the traditional Chinese medicine composition has no obvious toxicity to main organs such as heart, spleen, lung, kidney and the like, and shows good safety and clinical transformation potential.

The application belongs to the field of microorganism eukaryotes, and the strain preservation date is August 18, 2025, and the preservation number is CGMCC NO.35650. The vaccine is prepared by isolating and inactivating the Meyerozyma guiliardmi yeast ML001, the vaccine can significantly reduce the alcohol-induced ALT and AST levels in mice, reduce liver lipid accumulation, down-regulate fatty acid synthesis genes Fasn and Acly and up-regulate fatty acid oxidation genes Ppara and Cpt1a , thereby effectively inhibiting lipid synthesis and activating fatty acid oxidation, reducing alcoholic liver injury, and having no obvious toxicity to main organs such as heart, spleen, lung and kidney, and showing good safety and clinical transformation potential.

The present invention provides an RNA framework and a gene editing method for gene editing. The RNA framework comprises an upstream sequence of target site, a sequence to be inserted and a downstream sequence of target site along the direction of 5′→3′. Based on the intrinsic mechanism of eukaryotes, this gene editing method uses RNP or RNA (which can be prepared in vitro) and related proteins as vectors to transfer into the cytoplasm and nucleus to achieve gene editing of specific sequences or loci on the genome of the target system, such as insertion, deletion, replacement of specific sequences and site replacement, and has high targeting accuracy. The present invention is more suitable for further clinical application than other prior art because it does not introduce exogenous systems or substances such as proteins derived from prokaryotes and does not produce double-strand breaks.

It has been established that one or more large double stranded DNA fragments (each 2,000 to 40,000 base pairs in size) can be captured and isolated from genomic DNA fragments using sequence specific PNA hybridization probes. Compositions and methods for enrichment of a multiplicity of long DNA sequences selected from the genome of any eukaryote are provided. Capture is performed using multiple PNA molecules with gamma-modified chiral backbones, comprising a mixture of neutral and positive chemical groups. Two or more PNA probes with covalently bound haptens, preferably biotin, target each DNA domain of interest for capture, isolation, and subsequent sequencing analysis of the multiplicity of enriched targets, including DNA methylation sequencing. The methods include enhancement of probe-DNA binding specificity through single strand binding proteins (SSB).

The invention relates to an AnaCas9 mutant as well as a gene editing system and application thereof, and relates to the technical field of biology. The AnaCas9 mutant is modified at a specific site, so that the gene editing activity of the AnaCas9 mutant is remarkably improved, and the AnaCas9 mutant can be widely applied to efficient gene editing of prokaryotes and eukaryotes and particularly can also realize efficient gene editing in mammalian cells (including human cells) and saccharomycetes.

Provided herein is technology relating to nucleotide repeat elements present in the genomes of eukaryotes and particularly, but not exclusively, to technologies for reducing the levels of disease-causing products expressed from expansions of nucleotide repeats.