248 results about "Ribosome" patented technology

The invention provides an UTR (Untranslated Region) element H2202 P1-G as well as a construction method and application thereof, and relates to the technical field of mRNA (messenger ribonucleic acid). According to the present invention, the ribosome load prediction and the secondary structure optimization are performed on the natural 5 'UTR of the HIV TAT 202 gene through the BaidleHelix platform, and the obtained HTAT 202 P1 sequence avoids the inhibitory hairpin structure so as to significantly improve the luciferase expression quantity compared to the natural UTR; an ncRNA sequence without a secondary structure is introduced on the basis of the HTAT 202 P1, translation inhibition of a 5 'cap region is further relieved, and the protein expression quantity of the constructed H2202 P1-G mutant (the DNA sequence of the H2202 P1-G is as shown in SEQ NO 1, and the RNA sequence is as shown in SEQ NO 2) is further improved.

The invention provides an UTR element NHP1 as well as a construction method and application thereof, and relates to the technical field of mRNA. A 5 'UTR with a good expression effect is designed by integrating dominant sequences of a human high-expression gene and a pathogen natural UTR, a chimeric structure NHP1 with high ribosome load is predicted through a calculation model, a DNA sequence of the NHP1 is as shown in SEQ NO 1, and an RNA sequence of the NHP1 is as shown in SEQ NO 2; an EGFP report system is adopted on the DNA level to rapidly screen UTR; the translation efficiency is quantitatively evaluated on the RNA level through luciferase mRNA (N1-methyl pseudouridine modification); and the particle size is controlled by a microfluidic technology, so that the optimized UTR-mRNA is efficiently expressed after being delivered.

This application relates to the field of biosynthesis technology, and specifically discloses a pyridone compound, its biosynthesis method and application. Based on synthetic biology-related technologies, this application modifies non-ribosomal peptide synthetase, heterologously expresses non-ribosomal peptide synthetase mutants in a variety of hosts, and for the first time discovers and obtains a pyridone compound, and determines its chemical formula as C5H4N2O3, named 5-amino-pyridine-2,3,6-trione or 5-hydroxy-3-imino-pyridine-2,6-dione. This novel pyridone compound can be used as a raw material or intermediate in the development and preparation of novel antibiotics, drugs and dyes.

The invention discloses a combined module for improving the yield of actinomycetes polyketone natural products and construction and application of recombinant bacteria containing the combined module, and belongs to the technical field of genetic engineering. In order to improve the yield of natural products such as polyketone, non-ribosome peptide and the like, the invention provides a combined module capable of improving the yield of actinomycetes natural products by combining and transforming a synthesis pathway of CoASH and a post-modification pathway of PPTase through a metabolic engineering strategy, and the module consists of CoASH cofactor synthesis module elements SCoaAR106A and SCoaD and a PPTase post-modification module element HPC3. By constructing a recombinant vector and recombinant bacteria containing the combined module, the yield of natural products such as actinomycetes polyketone, polyether and non-ribosomal peptide can be effectively improved by over-expression of the combined module.

The invention provides a lysine artificial operon and application thereof. The gene sequence of the artificial operon comprises a promoter, a lysC gene for coding aspartate kinase, an asd gene for coding aspartate semialdehyde dehydrogenase, a dapA gene for coding dihydropicolinic acid synthetase and a dapB gene for coding dihydropicolinic acid reductase, the ddh gene is used for coding diaminopimelate dehydrogenase; the lysA gene is used for coding diaminopimelate decarboxylase; except for the first lysC gene, sequences of RBS ribose binding sites of other genes are RBS ribose binding sites translated after gene transcription by intercepting 20-26nt DNA sequences at the upstream of a coding region ATG (adenosine triphosphate); the RBS ribose binding site is connected in front of the corresponding gene.

The invention relates to a brand-new microbial natural metabolite, namely Fcl-8, which is derived from a fermentation solution of Xenorhabdius budapestensis XBD8, and is characterized in that the Fcl-8 is a new microbial natural metabolite, namely Fcl-8, which is derived from the fermentation solution of the Xenorhabdius budapestensis XBD8. The purified Fcl-8 is a snow white flocculent crystal and is very easy to dissolve in water, and the solid state of the purified Fcl-8 is as shown in Figure 1. The chemical structural formula of the Fcl-8 is as shown in a figure 2, the chemical formula is C57H102N14O12, the molecular weight is 1175.78744, and the typical secondary mass spectrum information of the Fcl-8 is as shown in a figure 3. The Fcl-8 is derived from a microbial natural metabolite, is novel in structure and high in biological activity, and has very high potential to be developed into a novel biopesticide.

The invention relates to dsRNA of miscellaneous migratory locust ribosome large subunit protein genes RPL7 and RPL44 and application of the dsRNA, the dsRNA for synthesizing the miscellaneous migratory locust ribosome large subunit protein genes RPL7 and RPL44 is injected into miscellaneous migratory locust, the RPL7 and RPL44 genes of the miscellaneous migratory locust can be specifically silenced, and the miscellaneous migratory locust can be inhibited from feeding, so that the miscellaneous migratory locust loses nutrition supply and is abnormal in growth and development, and then individual death is caused; the method achieves the purpose of biological prevention and control, has the advantages of specificity, high efficiency and environmental protection on pest prevention and control, is a novel green and pollution-free locust prevention and control technology, and shows a huge application prospect on locust prevention and control.

The invention provides a macro virus group sequencing library construction method for removing host ribosome RNA through an enzyme method and a kit, and belongs to the technical field of biology. The method comprises the following steps: hybridizing total RNA (Ribonucleic Acid) of a sample with a DNA (Deoxyribose Nucleic Acid) probe aiming at host ribosome RNA to form a DNA / RNA hybrid chain; rNase H is used for digesting RNA in the hybrid chain, and DNase I is used for digesting the remaining DNA probe in the system; purifying to obtain virus RNA (Ribonucleic Acid); carrying out fragmentation treatment on the virus RNA; carrying out reverse transcription on the fragmented RNA to synthesize a first chain cDNA, then synthesizing a second chain cDNA, and carrying out terminal repair and dA tail addition to obtain double-chain cDNA; connecting the linker with the dT tail with the double-chain cDNA; and carrying out PCR amplification and purification on the connection product to obtain the macro virus group sequencing library. According to the construction method and the kit, the problem that the full-length sequence of the virus cannot be obtained from a low-load sample can be solved.

The invention is applicable to the technical field of biology, and provides an L7 / L12-PADRE sequence-multi-linked B cell epitope recombinant protein and a kit for detecting an anti-Brucella antibody. The recombinant protein is a multi-epitope tandem recombinant protein composed of Brucella ribosome L7 / L12 protein, a PADRE polypeptide sequence and multiple B cell epitopes, the amino acid sequence of the recombinant protein is as shown in SEQ ID No.1, and the recombinant protein is good in antigenicity. The anti-brucella antibody indirect ELISA (iELISA) detection method and kit established by taking the recombinant protein as the coating antigen have the characteristics of high sensitivity, strong specificity and good repeatability, are suitable for detecting the condition of generating the anti-brucella antibody by an organism, can clarify the immune background of brucella infection or brucellosis vaccine, and can be used for detecting the brucella infection or brucellosis vaccine. Basic data is provided for prevention and control of the Brucella disease of humans and animals, and meanwhile, the kit is conveniently applied to large-scale sample detection and epidemiological monitoring.

The invention discloses a saccharomyces cerevisiae recombinant strain based on a uracil metabolism gene URA3 as well as a construction method and application of the saccharomyces cerevisiae recombinant strain. The construction method comprises the following steps: taking saccharomyces cerevisiae rich in farnesyl pyrophosphate as an original strain, introducing a uracil metabolism gene URA3, synchronously overexpressing a ribosome biosynthetic gene UTP10 and an iron metabolism gene FIT3, and utilizing the uracil metabolism gene URA3 to express up-regulation correlation with the ribosome biosynthetic gene UTP10 and the iron metabolism gene FIT3, so as to obtain the farnesyl pyrophosphate-rich saccharomyces cerevisiae. The metabolic flux of the terpenoids is enhanced, and the saccharomyces cerevisiae recombinant strain for efficiently producing the terpenoids is constructed. According to the method, a metabolic network of yeast is systematically optimized, and combined regulation and control on precursor flux distribution, protein translation capability and metal ion steady state are realized, so that the microbial synthesis efficiency of sesquiterpenoids such as patchouli alcohol and beta-elemene is remarkably improved.

The invention provides a method for eliminating ribosomal RNA in a mouse total RNA sample, which comprises the following steps: carrying out reverse transcription on the total RNA in the mouse sample into cDNA, and amplifying the cDNA by adopting a primer sequence group to obtain a DNA fragment covering all sequences of the mouse ribosomal RNA; carrying out amplification by taking the DNA fragment as a template and adopting a single primer in the primer sequence group to obtain a single-stranded DNA probe; hybridizing the total RNA of the mouse sample by adopting a single-stranded DNA probe, and performing RNase H digestion to obtain a digestion product; sequentially carrying out DNase I digestion and RNA purification on the digestion product to obtain a target product; wherein the primer sequence group comprises upstream primers from the first to the nineteenth and downstream primers from the first to the nineteenth, and the primer sequence group has the characteristics of low cost, stable performance and high reduction efficiency.

The invention provides an oxazomycin high-yield engineering strain as well as a construction method and application thereof. The method for constructing the oxazomycin high-yield engineering strain comprises the following steps: driving high expression of a gene cluster by using a strong promoter in streptomyces longsheng, enhancing transcription of a key resistance gene module (ozmS-oxaA), and optimizing ribosome engineering of strain chassis cells and a culture medium. According to the invention, the rational synthetic biology and metabolic engineering technology is combined with the irrational ribosome engineering technology to gradually increase the yield of the streptomyces longsheng oxazomycin, and the established efficient biosynthesis system lays a foundation for further development of the oxazomycin.

This invention relates to a combined module for increasing the yield of polyketide natural products from actinomycetes, and the construction and application of recombinant bacteria containing this module, belonging to the field of genetic engineering technology. To improve the yield of natural products such as polyketides and nonribosomal peptides, this invention utilizes a metabolic engineering strategy to combine and modify the CoASH synthetic pathway and the PPTase post-modification pathway, providing a combined module that can increase the yield of natural products from actinomycetes. This module consists of the CoASH cofactor synthesis module element SCoaA. R106A Composed of SCoaD and PPTase post-modification module element HPC3, recombinant vectors and recombinant bacteria containing this module were constructed. It was found that overexpression of this module can effectively increase the yield of natural products such as actinomycete polyketides, polyethers, and non-ribosomal peptides.

The invention is applicable to the technical field of genetic engineering and biological medicine, and provides construction of a fusion recombinant protein ABT for preventing brucellosis and application of the fusion recombinant protein ABT in preparation of protective vaccines. The fusion recombinant protein ABT is composed of an immunological enhancement antigen and a multi-epitope tandem antigen, wherein the immunological enhancement antigen (named as A) comprises a Brucella ribosome L7 / L12 protein and a PADRE sequence; the multi-epitope tandem antigen is formed by connecting a dominant B cell epitope part (named as B), a dominant Tc cell epitope part and a dominant Th cell epitope part (named as T) in series, wherein the dominant B cell epitope part (named as B) is derived from Brucella SurA, OMP31, BP26 and Trigger factor proteins. The ABT Brucella multi-epitope subunit vaccine prepared by mixing the fusion recombinant protein ABT and an immunologic adjuvant has the characteristics of good purity, high safety and strong immunogenicity, can stimulate an organism to generate a protective antibody, and has long antibody maintenance time.

The invention provides a compound as shown in a formula (I), and a stereoisomer, a tautomer, an isotope label, nitrogen oxide, a solvate, a polymorphic substance, a metabolite, ester, pharmaceutically acceptable salt or prodrug thereof, or a pharmaceutical composition thereof, which have better antibacterial and anti-inflammatory effects, can be used as an antibiotic, and can be used for preparing a pharmaceutical preparation for preventing and treating diseases. The compound can treat infection caused by pathogenic microorganisms resistant to erythromycin and telithromycin, and simultaneously acts on double targets of ribosome and topoisomerase.

Provided are a non-ribosomal antifungal peptide MaAFP1, a preparation method therefor and a use thereof. The molecular formula of the antifungal peptide MaAFP1 is C41H74N18O12, wherein an amino acid sequence from the N-terminus to the C-terminus is (D)Arg-(L)Arg(OH)-(D)Orn-(L)Thr-(D)Orn-(L)Arg(OH)-(D)Tyr. Fermentation is performed by using Metarhizium anisopliae CQMa421, a fermentation broth is collected, and separation and purification are performed by means of ion exchange chromatography and high performance liquid chromatography so as to obtain the antifungal peptide MaAFP1. The structure of MaAFP1 is analyzed by comprehensively using an Edman degradation method, mass spectrometry, infrared spectroscopy, nuclear magnetic resonance spectroscopy, and circular dichroism spectroscopy. By performing an in vitro bacteriostatic test, it is found that MaAFP1 has a good inhibitory effect on a variety of animal and plant pathogenic fungi, and has the value of the development of a novel antibacterial drug.

The application discloses a LAMP primer group, a kit and a detection method for detecting Pinus armandii rust fungus, and belongs to the technical field of genetic engineering. The application takes the specific nucleotide sequence of the large subunit ribosomal RNA of the Pinus armandii rust fungus as a target gene to design and screen a primer group which is specific and high in sensitivity, and can be used for LAMP detection of the Pinus armandii rust fungus. The LAMP primer group provided by the application has the advantages of high universality, high specificity, high sensitivity and good accuracy, and the detection limit can reach 4.5x10 ‑4 ng / μL, which is 100 times higher than the sensitivity of the LAMP detection of different species in the same genus in the prior art, the detection method can rapidly detect the Pinus armandii rust fungus from spore piles and host plant materials, and is suitable for the detection of Pinus armandii seedling materials in forest fields and customs quarantine ports.

The present invention relates to the application of phosphopantetheinyltransferase E7X1, its encoding gene, recombinant vectors containing the gene, and recombinant bacteria, belonging to the field of genetic engineering technology. To increase the yield of natural products such as polyketides and non-ribosomal peptides, the present invention utilizes bioinformatics analysis and other means to mine a highly efficient phosphopantetheinyltransferase E7X1 from a database. The amino acid sequence of the enzyme is shown in SEQ ID NO.1, and the nucleotide sequence of the gene encoding the enzyme is shown in SEQ ID NO.2. By constructing a recombinant vector and recombinant bacteria containing the enzyme encoding gene, the present invention discovered that overexpressing the enzyme can effectively increase the yield of natural products such as polyketides, polyethers, and non-ribosomal peptides.

Provided are a method for preparing scar-free circular RNA, and a recombinant nucleic acid molecule for preparing scar-free circular RNA and use thereof. The provided recombinant nucleic acid molecule based on the IRES stem-loop structure can be used for preparing a circular nucleic acid molecule completely free of exogenous sequences without altering any sequence of the IRES, thereby avoiding unpredictable effects on ribosome recruitment function caused by IRES mutations.

A cassette for the delivery of one or more mRNA is provided, the cassette comprising a 5' UTR, a Nodamura replicon, a ribosomal skipping polynucleotide, the one or more mRNA, a plant virus assembly origin polynucleotide ("OAS"), and a 3' UTR. In one aspect, the cassette further comprises a 5'mRNA cap. In a yet further aspect, the cassette does not comprise a poly A tail.

This invention relates to the field of bioinformatics, and particularly to a method and system for denoising single-cell immune repertoire sequencing data. The method includes data preprocessing and feature extraction, bidirectional collaborative denoising, intelligent comprehensive judgment and classification, data archiving and background learning, and result output. Compared to existing technologies that primarily rely on static thresholds for cell filtering, which struggle to comprehensively assess and eliminate multi-dimensional noise, leading to incomplete purification and the potential deletion of high-value cell information, this invention employs a systematic denoising scheme integrating multi-parameter dynamic threshold filtering, specific gene contamination analysis, and targeted optimization of VDJ data. It sets dynamic thresholds by integrating multi-dimensional quality control indicators such as UMI number, gene number, and the proportion of mitochondrial and ribosomal genes, and specifically identifies and filters interfering genes and background sequences. This enables refined and hierarchical removal of complex noise, significantly improving the overall quality of cell datasets and the accuracy of VDJ receptor sequence analysis.

This invention discloses a *Bacillus laterosporus* strain for the stable production of nonribosomal cyclic lipolipeptide and its construction method. The M1A gene in the Bogorol biosynthetic gene cluster is knocked out using CRISPR-Cas9 technology to block the Bogorol pathway, which competes with Brevicidine for substrates. Subsequently, the pMCCas9-S48 plasmid, which integrates S8 and S41 sgRNAs and their homologous arms as repair templates, is used to simultaneously knock out S8 and S41 family peptidase genes, preventing the expression of extracellular degradation enzymes. This invention also provides the application of the above strain in the preparation of nonribosomal cyclic lipolipeptide of Brevicidine and a method for preparing the nonribosomal cyclic lipolipeptide. This strain, through reducing metabolic competition and preventing product degradation, exhibits rapid cell growth, stable production of Brevicidine, and significantly reduced degradation rate, resulting in a 66.13% increase in fermentation yield compared to the unmodified strain. This invention solves the problems of degradation and unstable yield in the production of Brevicidine, provides an effective technical approach for the industrial production of Brevicidine, and provides important technical support for the development and application of novel antimicrobial peptide drugs.

An immunogenic composition comprising: a) one or more Plasmodium-derived ribosomal or ribosomal associated protein or immunogenic fragment thereof which has a sequence which is at least about 80%, 85%, 90%, 95%, 98%, 99% or 100% identical to a ribosomal or ribosomal associated protein or an immunogenic fragment of a ribosomal or ribosomal associated protein recited in FIG. 1; or a ribosomal or ribosomal associated protein or peptide or immunogenic fragment thereof as recited in FIG. 2 or FIG. 3; and / or b) a polynucleotide encoding one or more protein, peptide or immunogenic fragment of a); wherein the immunogenic composition is for use in eliciting an immune response in a subject to treat or prevent malaria. Also provided are Plasmodium-derived ETRAMPs and / or histones, or immunogenic fragments thereof, for use in eliciting an immune response in a subject, preferably to treat or prevent malaria.

Provided herein are methods for detecting and / or measuring mRNA associated with ribosomes in a cell. The method can be used to assess potency and other characteristics of an miRNA drug product. Also provided are related products, including cells and reagents.

The present invention provides compositions and methods for inhibiting reverse transcription of ribosomal RNA. The composition for inhibiting reverse transcription of ribosomal RNA according to the present invention is characterized by containing a polymer having at least one functional group selected from the group consisting of a sulfuric acid group, a sulfo group, and salt-type groups thereof, and by being used in an amount such that the concentration of the polymer in a reverse transcription reaction solution reaches 10 pg / [mu] L or more.

A translation-independent method of using directed evolution to engineer aminoacyl-tRNA synthetases (aaRSs) that directly select for tRNA acylation without ribosomal translation (START) is described. The efficacy of START was demonstrated by identifying novel mutants of the M. alvus pyrrolysyl-tRNA synthetase from a naïve library that charge noncanonical amino acids.

The invention discloses an efficient ribosome imprinting sequencing library construction method, and belongs to the technical field of biology. In the enzyme digestion treatment step, a mixed enzyme system of RNase I and micrococcus nuclease is adopted to treat the cell lysis buffer, and the trinucleotide periodicity of RPFs can be remarkably improved to 80% or above. In the target fragment recovery step, the sucrose gradient ultracentrifugation step is omitted, the RPFs are directly recovered by gel electrophoresis, a plurality of enzyme reaction steps in library construction are optimized and integrated, and streptavidin magnetic beads are used for solid phase purification, so that the whole library construction period is greatly shortened from 7 days to 2.5 days. The method does not depend on special equipment such as ultracentrifugation, and can be completed in a conventional molecular biology laboratory. Besides, a multi-enzyme synergistic RNA terminal one-step modification system is innovatively constructed, single-tube one-step completion is realized, the terminal treatment efficiency and consistency are improved, and a key guarantee is provided for obtaining a high-quality and high-complexity Ribo-seq library.

The invention discloses construction of a shewanella hydrogengens-CdS nanoparticle hybrid system based on a hydArsp gene and an application of the shewanella hydrogengens-CdS nanoparticle hybrid system. The method comprises the following steps: connecting a hydArsp gene derived from Rhodobacter sphaeroids coded hydrogenase to a vector pYYDT, carrying out ribosome optimization transformation to obtain a plasmid pYYDT-hydArsp, introducing the recombinant plasmid pYYDT-hydArsp into wild shewanella oneidensis MR-1 to obtain a recombinant shewanella engineering bacterium SERS with high hydrogen production efficiency, anchoring CdS nanoparticles on periplasm and an extracellular membrane of a strain in a self-assembly manner, and carrying out high-yield hydrogen production on the recombinant shewanella engineering bacterium SERS. The CdS / SERS high-hydrogen-yield active hybrid system is obtained. The CdS / SERS hybrid system constructed by the invention endows shewanella with light capturing ability, CdS nanoparticles provide photoelectrons for engineering bacteria to promote intracellular reducing power and ATP regeneration, and the hybrid system constructed by the invention can enhance the proton reduction rate of the engineering bacteria and improve the hydrogen production efficiency.

Methods and kits of rapidly isolating molecules from a sample are provided. Methods of treating a subject in need thereof using the rapid isolation kits and / or methods described herein are also provided. In some embodiments, the molecules are selected from ribosomes, DNA, RNA, and protein. In some embodiments, the sample is selected from a tissue sample, a blood sample, a non-blood biological fluid sample, and a whole organism sample.

The present disclosure provides compositions, methods of making, and use of a circular ribonucleic acid (circRNA) therapeutic for targeted delivery to immune cells, including T cells, for effector functions such as B cell depletion, providing, among other things, methods of treating B cell-mediated diseases such as autoimmune disease and cancer, meeting the medical need for efficacious and safe therapies that overcome the disadvantages of ex vivo CAR T therapy. Provided by the present disclosure is a circRNA therapeutic comprising: (i) a circRNA comprising a ribosome recruiting element, and a coding sequence encoding a CAR comprising, for example, an anti-CD19 antibody or antigen-binding fragment thereof, and (ii) a targeting moiety (e.g., anti-CD5) that binds to an immune cell (e.g., a T cell), wherein the circRNA is in a delivery vehicle, and a targeting moiety that binds to a T cell is conjugated to the delivery vehicle (e.g., a lipid nanoparticle).