9667results about "Transferases" patented technology

The present disclosure relates to materials and methods for spatially analyzing nucleic acids that have been fragmented with a transposase enzyme, alone or in combination with other types of analytes.

The present disclosure relates to materials and methods for spatially analyzing nucleic acids that have been fragmented with a transposase enzyme, alone or in combination with other types of analytes.

The present disclosure relates to materials and methods for spatially analyzing nucleic acids that have been fragmented with a transposase enzyme, alone or in combination with other types of analytes.

The present disclosure provides methods and compositions for detecting and spatially locating analyte interactions and gene expression in a biological sample. For example, provided herein are methods of determining a location of at least one analyte in a biological sample using analyte-binding moieties, proximity ligation, and an array including capture probes.

The present disclosure relates to materials and methods for spatially analyzing nucleic acids that have been fragmented with a transposase enzyme, alone or in combination with other types of analytes.

The present disclosure relates to materials and methods for spatially analyzing nucleic acids that have been fragmented with a transposase enzyme, alone or in combination with other types of analytes.

The present disclosure relates to materials and methods for spatially analyzing nucleic acids that have been fragmented with a transposase enzyme, alone or in combination with other types of analytes.

The present disclosure relates to materials and methods for spatially analyzing nucleic acids that have been fragmented with a transposase enzyme, alone or in combination with other types of analytes.

Methods and systems are provided for generating and utilizing a genetically engineered bacterium comprising a modification in glnD, wherein said modification is selected from the group consisting of: deletion of the entire gene, deletion of substantially the entire gene, deletion of an ACT domain, deletion of more than 50% of an ACT domain, deactivation of an ACT domain, and deactivation of an UTase domain.

Systems for plant culture include a chamber featuring one or more walls enclosing a spatial volume internal to the chamber, where the one or more walls include a surface for supporting a plant within the enclosed spatial volume, a gas delivery apparatus with at least one gas source, a nutrient delivery apparatus with a reservoir, a sampling apparatus connected to a port formed in the one or more walls, and a controller configured so that during operation of the system, the controller activates the nutrient delivery apparatus to deliver an aqueous growth medium to the plant, and activates the gas delivery apparatus to deliver into the enclosed spatial volume a mixture of isotopically-substituted gases. Also provided are methods of use of the system for measuring nitrogen in a plant and for identifying microbes capable of providing fixed nitrogen to a plant.

The invention belongs to the technical field of biology, and particularly discloses a methyltransferase mutant high-throughput screening method, a methyltransferase mutant and application. The method can be used for high-throughput screening of methyltransferase mutants, the transformation efficiency of methyltransferase is improved, and high-activity methyltransferase mutants are obtained; the methyltransferase mutant provided by the invention can obviously improve the activity of methyltransferase, and is used for producing Nacetyl-5-methoxytryptamine to improve the yield. The activity of the methyltransferase can be remarkably improved by using the engineering bacterium expressing the methyltransferase mutant, the engineering bacterium is used for producing Nacetyl-5-methoxytryptamine to improve the yield, and the yield of the Nacetyl-5-methoxytryptamine is up to 22.7 mM.

Aspects of the disclosure relate to methods of reducing expression or activity of DMPK (e.g., reducing the level of a mutant or wild-type DMPK RNA, or the activity of a DMPK gene product) and / or methods of treating myotonic dystrophy (e.g., DM1) in a subject. In some embodiments, the methods comprise administering to the subject a composition comprising complexes (e.g., muscle targeting complexes) comprising an oligonucleotide (e.g., a DMPK—targeting oligonucleotide) covalently linked to an antibody (e.g., anti-TfR1 antibody).

The invention discloses a biosynthesis system for high-yield 1, 6-hexamethylenediamine and application of the biosynthesis system, and belongs to the technical field of biology. According to the method, key genes which are excavated and optimized in a 1, 6-hexamethylenediamine biosynthetic pathway are co-expressed in a single cell module or a multi-cell module, and different enzyme elements expressed by the cell module are subjected to enzyme cascade reaction, so that 1, 6-hexamethylenediamine can be efficiently synthesized by taking caprolactam or 6-aminocaproic acid as a substrate; furthermore, after the carboxylic acid reductase MaCAR is mutated, the catalytic activity of the carboxylic acid reductase MaCAR can be remarkably improved, and the yield of the 1, 6-hexamethylenediamine is further improved; in addition, the biosynthesis system disclosed by the invention directly takes the cell module as a whole-cell catalyst, and an enzyme purification step is avoided, so that the biosynthesis system has the advantage of low cost.

The invention discloses an acyltransferase mutant and application of the acyltransferase mutant in synthesis of N-acetyl-trans-4-hydroxyproline, and belongs to the technical field of enzyme engineering. The acyltransferase mutant provided by the invention contains 10th and 95th mutations on the basis of a corresponding amino acid sequence, such as a sequence as shown in SEQ ID NO.1, or contains 62nd or 64th mutations while containing the 10th and 95th mutations. Compared with an original enzyme before mutation, the acyltransferase mutant obtained by the invention has higher acyltransferase activity, and the conversion rate of trans-4-hydroxy-L-proline is greatly improved, so that the yield of N-acetyl-trans-4-hydroxy proline is further improved, and the yield of N-acetyl-trans-4-hydroxy proline is improved. After the surfactant and the metal ions are added, the yield of the N-acetyl-trans-4-hydroxyproline synthesized by the mutant D62K / D10V / D95I under the catalysis is up to 18.7 g / L.

The invention belongs to the technical field of RNA biological prevention and treatment, and particularly relates to dsRNA for preventing and treating crop anthracnose and application of the dsRNA. The dsRNA is targeted inositol phosphatidylceramide (IPC) synthetase coding gene dsRNA, and is obtained by transcription of a part of sequence (SEQ ID No.1) of an inositol phosphatidylceramide (IPC) synthetase coding gene CaAUR1, the gene segment is a segment of a target gene for preventing and treating crop anthracnose, and the dsRNA obtained by transcription can obviously reduce pathogenicity of mango anthracnose bacteria, and has the effects of keeping mango fresh, preventing and treating anthracnose. And the compound can be used for preventing and treating the crop anthracnose or preparing products for preventing and treating the crop anthracnose.

The invention discloses polyketide synthase or a polyketide synthase mutant and application thereof, and belongs to the technical field of gene engineering. The polyketide synthase is AspiPKS4, AspiPKS5, AspiPKS6 and AspiPKS7 from cyathea spinulosa, FhPKS2, FhPKS3, FhPKS4, FhPKS5 and FhPKS6 from ficus himalayana, and CrPKS from floating fern. The mutant is obtained by carrying out single point mutation on the second amino acid of AspiPKS7 or deleting an N-terminal redundant sequence. The polyketide synthase or the mutant is used for replacing NnHisspS in an original FBP luminescence system, and the biological self-luminescence level can be remarkably improved. The invention further provides a simplified and improved FBP system which comprises the polyketide synthase or the polyketide synthase mutant, CPH, H3H and Luz, and the application convenience and the application luminescence capacity of the FBP system are improved.

The invention discloses a glycosyl transferase UGT76G1 mutant and a method for catalytically synthesizing rebaudioside M by using the glycosyl transferase UGT76G1 mutant, and relates to the technical field of biological catalytic synthesis. The glycosyl transferase UGT76G1 mutant is any one of the following components (A)-(C): (A) a protein obtained by performing any one or more of the following mutations on the basis of an amino acid sequence as shown in SEQ ID NO.1: the 89th amino acid is mutated into H from M; the 380th amino acid is mutated from L to M; the 411th amino acid is mutated from A to Y; (B) a protein which has 95% or 98% or more of identity with the amino acid sequence defined by (A) and has the same function as the amino acid sequence defined by (A); and (C) a fusion protein obtained by connecting a tag to the terminal of the protein defined in (A) or (B). The glycosyl transferase UGT76G1 mutant disclosed by the invention is obtained by mutating and screening glycosyl transferase UGT76G1, has higher catalytic activity, and can be used for efficiently synthesizing rebaudioside M by taking rebaudioside D and UDPG as substrates.

The invention discloses a radish bolting and flowering gene RsSUF4 regulated by m6A demethylase and application of the radish bolting and flowering gene RsSUF4, relates to the technical field of bioengineering, and is technically characterized in that the gene is RsSUF4. The functions of the demethylase RsALKBH10B and the flowering inhibition gene RsSUF4 in the flowering process are analyzed in a targeted manner, and a specific association mechanism between m6A modification and radish flowering regulation and control is defined.

The invention provides recombinant escherichia coli with high yield of ergothioneine and a method for preparing ergothioneine, the recombinant escherichia coli co-expresses ergothioneine anabolic enzymes Egt1 and Egt2, the methyltransferase structural domain of the Egt1 is derived from trichoderma reesei, mycobacterium smegmatis EgtD or rhizopus nigricans, and the methyltransferase structural domain of the Egt2 is derived from the trichoderma reesei, the mycobacterium smegmatis EgtD or the rhizopus nigricans. The histidine sulfoxide synthase structural domain is derived from trichoderma reesei, schizosaccharomyces pombe or rhizopus nigricans, and the Egt2 is derived from claviceps purpurea, neurospora crassa or aspergillus niger. After the recombinant escherichia coli obtained by screening is cultured for 68 hours in a 5L fermentation tank, the yield of ergothioneine can reach 5g / L or above; wherein after recombinant escherichia coli co-expressing Egt1 enzyme (the structural domain of methyltransferase is derived from mycobacterium smegmatis EgtD, and the structural domain of histidine sulfoxide synthase is derived from trichoderma reesei) and Egt2 enzyme (derived from neurospora crassa) are cultured in a 5L fermentation tank for 68 hours, the yield of ergothioneine can reach 8.7 g / L, and the production intensity reaches 127.9 mg / (L.h).

The present disclosure provides compositions and methods for prime editing with improved editing efficiency and / or reduced indel formation with modified prime editors and prime editor fusion proteins. The disclosure further provides, vectors, cells, and kits comprising the compositions and polynucleotides of the disclosure.

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 discloses a glycosyl transferase mutant and an application of the glycosyl transferase mutant in synthesis of rebaudioside. According to the invention, single-point mutation and multi-point mutation are carried out on the basis of a glycosyl transferase amino acid sequence as shown in SEQ ID NO: 1, and a mutant with improved catalytic activity is obtained. The catalytic activity, the substrate specificity and / or the substrate specificity of the glycosyl transferase mutant are / is changed, and the catalytic activity of enzyme to a specific substrate can be remarkably improved by mutation at a specific site. And carrying out induced expression and protein purification on the obtained mutation sequence to obtain the mutant enzyme. The mutant enzyme is used as a catalyst, and UDPG is used as a glycosyl donor, so that the catalytic reaction efficiency of substrates such as stevioside ST, rebaudioside A (RebA) and rebaudioside D (RebD) can be obviously improved. According to the glycosyl transferase UGT76G1 mutant constructed by the research, the catalytic activity of the glycosyl transferase UGT76G1 mutant is improved, and efficient production of rebaudioside M is realized by optimizing a reaction system.

The invention relates to the technical field of genetic engineering, and particularly discloses a genetic engineering bacterium for producing melanin as well as a construction method and application of the genetic engineering bacterium. According to the application, the yield of the melanin can be effectively increased by modifying a metabolic pathway of the genetically engineered bacterium, and the yield of the melanin reaches 6.2 g / L and 18.5 g / L respectively through shake-flask culture and 5L fermentation tank culture. Compared with the prior art, the yield of melanin produced by the genetically engineered bacterium is improved by 30% at the shake flask fermentation level, the fermentation yield of a fermentation tank is improved by 4.3 times, in addition, expensive tyrosine is changed into glucose as a fermentation precursor substance, the production cost is greatly reduced, and an efficient and feasible solution is provided for large-scale production of melanin.

The invention discloses a mesophilic carotenoid-producing recombinant yarrowia lipolytica strain as well as a construction method and application thereof, the recombinant strain exogenously expresses mesophilic original fatty acid desaturase (FAD3) and isocitrate dehydrogenase (IDH2) mutants from phaffia rhodozyma, and efficient synthesis under the condition of 30 DEG C is realized; secondly, a metabolic pathway of the recombinant strain is optimized based on a metabolic flow push-pull strategy, and isopentenyl pyrophosphate isomerase (IDI), farnesyl pyrophosphate synthase (ERG20), plant alkene dehydrogenase (CarB) and bifunctional plant alkene synthase (CarRP) are overexpressed, so that the flux of a precursor is increased; finally, gene multi-copy of astaxanthin synthase (crtS) and cytochrome P450 reductase (crtR) from phaffia rhodozyma is carried out, and the carotenoid production capacity of yarrowia lipolytica is further improved. The construction method of the mesophilic yarrowia lipolytica is simple, efficient synthesis of carotenoid can be better achieved at the higher temperature, the fermentation cost is greatly reduced, and the industrial production potential is achieved.

The invention discloses an attenuated salmonella typhimurium EN-VNP (Enhanced Natriuretic Peptide) strain as well as a preparation method and application of the attenuated salmonella typhimurium EN-VNP strain. Comprising the following steps: on the basis of a salmonella VNP20009 strain, flagellin genes fliC and fljB, a phosphoethanolamine transferase gene eptA, a 4-amino-4-deoxy-L-arabinose transferase gene arnT and phosphotransferase yeiU are sequentially knocked out or silenced, and a strain is formed after a phosphotransferase gene lpxE is knocked in or overexpressed, so that the attenuated salmonella typhimurium EN-VNP strain is prepared. According to the attenuated salmonella typhimurium EN-VNP with low toxicity and the T cell adjuvant effect and an EN-OMV antigen delivery system prepared from the attenuated salmonella typhimurium EN-VNP, an improved strain does not contain flagellin, an LPS molecular structure is changed, the attenuated salmonella typhimurium EN-VNP has the low toxicity and the T cell adjuvant effect, and normal multiplication culture can be achieved.

The invention discloses N-acetylglucosamine transferase AcetRL-R349H and an application thereof, and belongs to the technical field of functional enzymes. The amino acid sequence of the N-acetylglucosamine transferase AcetRL-R349H is as shown in SEQ ID NO. 5 (sequence identifier number 5). The invention further discloses the application of the N-acetylglucosamine transferase AcetRL-R349H in the preparation of the chitosan oligosaccharide. The N-acetylglucosamine transferase AcetRL-R349H disclosed by the invention is obtained by carrying out mutation modification on the N-acetylglucosamine transferase AcetRL, and can be used for producing the chitosan oligosaccharide with the polymerization degree of 4 to 6, especially (GlcNAc) 6. The product spectrum of the N-acetylglucosamine transferase is widened, and the N-acetylglucosamine transferase can be used for preparing high-polymerization-degree chitosan oligosaccharide and has a wide application prospect.

The invention belongs to the field of gene engineering, and particularly relates to application of an XC3225 gene in increasing the yield of xanthomonas campestris pv. Campestris xanthan gum. The invention finds that the XC3225 gene has a negative regulation effect on the xanthan gum yield, and the xanthan gum yield is remarkably improved by inhibiting or knocking out the XC3225 gene in the genome of xanthomonas campestris pv. Campestris, so that a new gene resource is provided for xanthan gum preparation.

The invention discloses a hemoglobin-resistant Taq DNA polymerase mutant and a construction method thereof, and relates to the field of biology, and the hemoglobin-resistant Taq DNA polymerase mutant is characterized in that a nucleotide sequence for coding the Taq DNA polymerase is shown as SEQ ID NO.1, and the mutant comprises at least one mutation or all mutations selected from S623D and E721A sites. On the basis of natural Taq DNA polymerase, the molecular structure of the Taq DNA polymerase is modified through rational design and combination with a site-specific mutagenesis biotechnology, so that the Taq DNA polymerase mutant is more suitable for PCR amplification of samples containing hemoglobin and the like than a wild type, and the result judgment accuracy of a sample containing a hemoglobin inhibitor is more facilitated.

The invention provides an RNA polymerase variant, and relates to the technical field of biology. The RNA polymerase variant provided by the invention can be used for remarkably reducing the generation of dsRNA impurities in an in-vitro transcription process and remarkably improving the integrity of an RNA transcription product, especially in transcription of long-fragment RNA. In addition, the invention provides an application of the RNA polymerase variant in in-vitro transcription, and also provides a method for preparing ultra-long saRNA (greater than 10000 nt).

The invention provides an application of an MET14 gene or an MET14 protein in improving the freezing resistance of saccharomyces cerevisiae. It is found for the first time that through overexpression of the MET14 gene or the MET14 protein, the freezing resistance of the saccharomyces cerevisiae can be remarkably improved, the saccharomyces cerevisiae can normally ferment and grow in frozen products, the excellent physiological activity of the saccharomyces cerevisiae in the frozen products is fully exerted, and then the quality of the frozen products is remarkably improved.