310 results about "Ribosome" patented technology

The invention relates to the technical field of biosynthesis, and particularly discloses a pyridone compound as well as a biosynthesis method and application thereof. On the basis of related technologies of synthetic biology, non-ribosome peptide synthetase is modified, non-ribosome peptide synthetase mutants are heterologously expressed in various hosts, a pyridone compound is found and obtained for the first time, the chemical molecular formula of the pyridone compound is determined to be C5H4N2O3, and the pyridone compound is named as 5-aminopyridine-2, 3, 6-triketone or 5-hydroxy-3-iminopyridine-2, 3, 6-triketone or 5-hydroxy-3-iminopyridine-2, 3, 6-triketone or 5-hydroxy-3-iminopyridine-2, 3, 6-triketone. The novel pyridone compound can be used as a raw material or an intermediate to be applied to development and preparation of novel antibiotics, drugs and dyes.

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 present invention relates to strategies for combining elements of non-ribosomal peptide synthase (NRPS) and polyketide synthase (PKS) into functional enzymes to promote the production of peptides and polyketide of any sequence. The present invention is based on the use of conserved sequence motifs within the T domain (or T domain-like) gene of the NRPS / PKS, which is used according to the invention as fusion sites for the production of new (and artificial) NRPS / PKS enzymes that can produce any peptide or polyketones of interest. The present invention provides artificial NRPS / PKS of the invention, methods and means for generating them, and nucleic acid libraries encoding various modules useful for designing NRPS / PKS for any peptide sequence of interest.

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 relates to an mRNA sequence ribosome occupancy prediction method, a model training method and a related device. The method comprises the following steps: acquiring an mRNA sequence and a secondary structure of the mRNA sequence in a target cell type; inputting the mRNA sequence and the secondary structure thereof into a pre-trained prediction model, respectively obtaining a first embedded representation of the mRNA sequence and a second embedded representation of the secondary structure through different encoders in the prediction model, fusing the first embedded representation and the second embedded representation based on a multi-head self-attention mechanism to obtain a fused embedded representation, and obtaining a multi-head self-attention prediction model; classification is carried out based on the fused embedded representation; and based on classified output of the prediction model, obtaining a prediction result of the ribosome occupancy of the mRNA sequence in the target cell type. Compared with the prior art, after the prediction model is trained in any cell type, ribosome occupancy prediction is carried out on any other cell type, the prediction precision is high, and the prediction model is high in mobility.

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 provides a construction method of recombinant escherichia coli for synthesis of ergothioneine, a strain and application of the strain, and relates to the technical field of construction of strains related to synthesis of ergothioneine, an NcEgt1 * sequence and an Egt D * sequence are fused and connected with a ribosome binding site sequence, and after XbaI and NotI enzyme digestion, the mixture is connected to a carrier pET-Duet-1 to obtain a recombinant plasmid pET-Duet-1-NcEgt1 *-EgtD *; the EgtE is connected to a recombinant plasmid pET-Duet-1-NcEgt1 *-EgtD * through Nd < e > I / KpnI enzyme digestion, and finally a recombinant vector pET-Duet-1-NcEgt1 *-EgtD *-EgtE is obtained; the method comprises the following steps: transforming a recombinant vector pET-Duet-1-NcEgt1 *-EgtD *-EgtE into E.coli BL21 to obtain a bacterial strain for producing ergothioneine, and naming the bacterial strain as E.coli Egt116; then, the recombinant plasmid pRSFDuet-1-EgtD *-EgtE is converted into a bacterial strain E.coli Egt116, and a bacterial strain E.coli Egt117 capable of producing ergothioneine is obtained; and finally, constructing a mutant strain of double genes of the strain Ecoli Egt117sdaA / MetJ by using a Cre-lox system, naming the mutant strain as E.coli Egt118, and increasing the ergothioneine yield of the strain to 218.71% in comparison with the ergothioneine yield of the strain E.coli Egt116. The recombinant strain constructed by the invention can be used for mass production of the ergothioneine, and large-scale production of the ergothioneine is realized.

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.

A method of treating Alcohol Use Disorder (AUD), Substance Use Disorder (SUD), tobacco use, pain, or proinflammatory disorders comprising: providing a subject with an effective amount of a modified tetracycline or derivative thereof to ameliorate or eliminate the AUD, SUD, tobacco use, pain, or proinflammatory disorder, and wherein the modified tetracycline or derivative thereof has reduced binding to a microbial ribosome and has the formula:wherein R1 is acetyl, R2 is OH or acetyl, R3 is acetyl, R4 is H or acetyl, and R5 is acetyl.

The invention discloses an OsRNE gene and application of protein coded by the OsRNE gene in rice leaf color regulation and chloroplast development. The nucleotide sequence of the gene OsRNE is as shown in SEQ ID NO.1, and the amino acid sequence of the encoded protein of the gene OsRNE is as shown in SEQ ID NO.2. According to the invention, the OsRNE is knocked out by using a CRISPR / Cas9 gene editing technology, and a functional loss mutant is obtained. The mutation of the gene causes damaged development of rice chloroplast, influences chlorophyll synthesis, albinism of seedlings and death within the trefoil stage, so that the gene can be used as a marker character in the seedling stage to assist rice molecular breeding. The OsRNE gene is mainly expressed in leaves, protein coded by the OsRNE gene is positioned in chloroplast, and development of the chloroplast and normal growth of seedlings are further affected by affecting metabolism of ribosome RNA in the chloroplast. Therefore, the method can be applied to rice leaf color character molecular genetic breeding, and has important significance in further understanding a rice chloroplast development regulation mechanism and improving the rice yield.

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.

Provided herein are nucleic acid inhibitors or activators that impact the expression, levels or activity of SNORA13 in a cell. Also provided are pharmaceutical compositions for using these activators and inhibitors for the treatment of ribosomopathies, cancer, and senescence-associated pathologies.

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 discloses a method for improving soluble expression of non-ribosome peptide synthetase in escherichia coli. According to the method, a polyion oligopeptide label is connected to the N end or the C end of the non-ribosome peptide synthetase; the method comprises the following steps: adding a polyion oligopeptide tag (D) 5 at the N / C end of ebony in a fusion protein SGE formed by non-ribosomal peptide synthetase ebony for producing decarboxylated carnosine and phosphokinase Sfp through a connecting peptide GSG, or replacing a GSG sequence in an original SGE sequence with the polyion oligopeptide tag (D) 5 to construct new plasmids, and transforming the obtained three new plasmids in recombinant escherichia coli to obtain the decarboxylated carnosine-producing non-ribosomal peptide synthetase ebony-phosphokinase Sfp. The solubility of the non-ribosome peptide synthetase in escherichia coli is greatly improved, and the addition of the polyion oligopeptide tag (D) 5 at the N end is more beneficial to reducing the formation of inclusion bodies and improving the yield of decarboxylated carnosine.

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.

The invention discloses a corn kernel development regulation gene ZmRH51 as well as an encoding protein, a functional marker and an application of the corn kernel development regulation gene ZmRH51. According to the application, a corn kernel mutant m223 is taken as a basic material, heterozygote of the corn kernel mutant m223 is hybridized with Zheng 58 (Z58) to construct an F2 segregation population, then phenotype identification and cytological observation are performed on the segregation population of the offspring, the gene ZmRH51 for controlling the mutation phenotype of the m223 is obtained through genetic analysis, BSR-seq analysis and map-based cloning, the gene is positioned in a nucleolus region, and the mutation phenotype of the m223 is controlled. The ZmRH51 is used for coding RNA helicase of a DEAD-Box gene family, and research shows that ZmRH51 participates in ribosome assembly and can influence processing of pre-rRNA in corn kernels; the excellent allelic variation genotype of the excavated ZmRH51 gene in the aspect of grain thickness is obtained through further excavation, the grain thickness and hundred-grain weight of the ZmRH51 gene can be increased through the utilization of the gene, and then the yield is increased. In addition, distribution and utilization potentials of different haplotypes in a selfing line are researched, and technical support and excellent germplasm resources are provided for utilization of the gene.

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.

The invention discloses phosphopantetheinyl transferase E7X1, a coding gene thereof, a recombinant vector containing the gene and application of recombinant bacteria, and belongs to the technical field of gene engineering. In order to improve the yield of natural products such as polyketones and non-ribosome peptides, the efficient phosphopantetheinyl transferase E7X1 is excavated from a database by means of bioinformatics analysis and the like, the amino acid sequence of the transferase E7X1 is as shown in SEQ ID NO.1, and the nucleotide sequence of the encoding gene of the transferase E7X1 is as shown in SEQ ID NO.2. By constructing a recombinant vector and recombinant bacteria containing the encoding gene of the enzyme, the yield of natural products such as polyketone, polyether, non-ribosome peptide and the like can be effectively increased by over-expressing the enzyme.

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.