189 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.

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 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.

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.

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.

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).

The use of different primer sets in reverse transcription incorporates tags allowing for the selective amplification of cDNA transcribed using the different primers. Primers targeting non-desired RNA sequences such as ribosomal RNA can be used to prevent subsequent amplification of cDNA transcribed from those non-coding fragments. Accordingly effective depletion of non-desired sequences after cDNA amplification can be achieved. Systems and methods of the invention have applications in whole-transcriptome analysis. Non-coding sequence targeting primers can include nucleotide analogs that, when enzymatically processed, prevent subsequent amplification. Library preparation can include single primer isothermal amplification (SPIA) techniques wherein an RNA sequence required for SPIA is included in random primers but is absent from primers targeting non-coding RNA.

The present disclosure relates to methods, devices and systems for exposing microorganisms or compositions comprising functional proteins to an oscillating electric field substantially free of a magnetic component in order to improve stress responses, survival and metabolite or protein production. In embodiments, a controller drives one or more insulated emitters with a true AC signal or pulsed DC signal, in stepped, swept or shuffled patterns of an electric field oscillating at one or more frequencies in the range 210 Hz to 99 kHz. Exposure to the field provides surmounting energy that enhances protein breathing, catalysis, and fundamental functions of multiprotein complexes including transcription factors and ribosomes, thereby improving stress resistance of microorganisms, their viability during fermentation, drying, storage, transport, and / or increasing production of functional or therapeutic proteins, metabolites or microbial lysates. The technology is applicable to manufacture of probiotics, therapeutics, metabolites, food, alcohol, feed and bioenergy, cosmetics, nutraceuticals and biopharmaceuticals..

The invention provides a preparation method for preparing a scar-free circular RNA (Ribonucleic Acid), a recombinant nucleic acid molecule for preparing the scar-free circular RNA and application of the recombinant nucleic acid molecule. The recombinant nucleic acid molecule based on the IRES stem-loop structure provided by the invention can be used for preparing a circular nucleic acid molecule in which an exogenous sequence is completely eliminated, and any sequence of IRES is not changed, so that the unpredictable influence of IRES mutation on recruitment of ribosome functions is avoided.

The invention discloses a genetically engineered bacterium for producing D-pantothenic acid based on a corynebacterium glutamicum CPSPC system as well as a construction method and application of the genetically engineered bacterium. Wild type C.glutamicum ATCC 13032 is taken as a chassis strain, and a functional combination of a glycine ribosome switch BsGR from bacillus subtilis and a promoter Ptuf / Ph36 is verified through fluorescent protein mCherry; a GCS system of escherichia coli and bacillus subtilis is introduced to the strain DPAg-15, genes related to synthesis of 5, 10-dimethyltetrahydrofolic acid are tested, it is found that serA delta197GTG / ATG significantly increases the yield of D-pantothenic acid, the yield of D-pantothenic acid of the constructed strain DPAj-2-tuf is increased to 23.06 g / L (63 h), the yield is 0.37 g / L / h, and the conversion rate is 0.17 g / g; the engineering strain DPAj-2-tuf is obtained by dynamically regulating and controlling a D-pantothenic acid synthesis path through a CPSPC system and combining plasmid overexpression serA delta197GTG / ATG, the yield of the engineering strain DPAj-2-tuf is increased by 2.1 times compared with that of a traditional method, and an efficient technical scheme is provided for producing D-pantothenic acid through a microbial fermentation method.

In the field of gene therapy, a major hurdle is the design and identification of constructs and gene therapy vectors providing therapeutic effects while displaying satisfactory safety profiles. In the treatment of Diamond-Blackfan Anemia (DBA), therapies alleviating several crucial anemia symptoms, such as blood or bone marrow cellularity, hemoglobin levels, erythrocytes levels, or platelet levels, while showing satisfactory safety profiles remain a challenge. The present invention provides constructs encoding ribosomal protein genes involved in DBA, such as genes encoding RPS19, RPS17, RPS24, RPS10, RPL35a, RPL11, RPS26, and RPL5, vectors, methods, cells, and medical uses thereof, addressing these challenges and finding particular applications in the field of autologous cell therapy treatment of DBA. Further, the present invention provides a non-genotoxic conditioning protocol for preparing a subject prior to cell therapy treatment for DBA using construct of the present invention.

Nucleic acid molecules comprising at least one transcription regulatory element operably linked to an open reading frame, wherein the open reading frame encodes a single RNA transcript encoding GCaMP7f, a ribosomal skipping peptide, and a fusion protein of a nuclear protein and photoactivatable red fluorescent protein are provided. Expression vectors and cells comprising the nucleic acid molecules are also provided, as are methods of using the nucleic acid molecules for simultaneous labeling and measuring calcium and analyzing a target cell.

The present invention relates to an oligonucleotide comprising: a) a nucleotide sequence of a broom-kojin-ricin loop (SRL) of a eukaryotic / mammalian 60S ribosomal subunit wherein the SRL nucleotide sequence comprises at least one adenine; and b) at least one cut-dependent marker; wherein the oligonucleotide is a single strand. Preferably, the oligonucleotides form at least one loop structure or stem-loop structure. In another aspect, the invention relates to a method of detecting active shiga toxin in a sample, the method comprising the steps of: a) providing at least one oligonucleotide according to the invention; b) providing a sample of shiga toxin to be detected; c) incubating the sample with at least one single-stranded oligonucleotide; and d) detecting a labeled signal, wherein the signal indicates the presence of Shiga toxin in the sample. The invention additionally comprises a kit comprising at least one oligonucleotide according to the invention.

This invention discloses a PhlD gene based on translation rate optimization and its applications, belonging to the fields of genetic engineering and protein engineering. Without altering the amino acid sequence of the PhlD protein, this invention optimizes the translation rate of the original PhlD sequence through synonymous codon substitution, enabling rapid translation within the coding domains. Translation deceleration sites are introduced into the contiguous regions of the coding sequence domains, matching the ribosomal translation rate with the protein co-translational folding process, significantly improving the soluble expression level of the PhlD protein. The PhlD gene with optimized translation rate results in a 5.5-fold increase in the soluble expression level of the encoded PhlD protein compared to the original sequence. Engineered bacteria containing this optimized gene show a 4-fold increase in the yield of phloroglucinol during fermentation compared to the control. This provides a core gene resource for the efficient soluble expression of the PhlD protein and the industrial production of phloroglucinol.

The invention discloses a high-yield cell-free protein synthesis system based on a cell crude extract and application of the high-yield cell-free protein synthesis system. The system comprises an escherichia coli extract, a DNA template, a buffer solution, an amino acid mixture, a nucleoside triphosphate mixture, an ATP regeneration system and antibiotics, the antibiotics are selected from compounds having inhibitory activity on metabolic processes except ribosome functions in prokaryotic microorganisms, including but not limited to one or more combinations of interfering DNA replication, DNA transcription, cell wall synthesis, energy metabolism, lipid synthesis or other non-translational related metabolic pathways. According to the method disclosed by the invention, the escherichia coli crude extract is used as a bacterial chassis, and non-essential metabolic pathways in a CFPS system are selectively inhibited by introducing different types of antibiotics, so that the yield of protein synthesis is increased.