8629results about "Transferases" patented technology

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.

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 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 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 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 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 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.

The invention provides a recombinant strain for controllably synthesizing high-proportion 4-hydroxybutyric acid (4HB) polyhydroxyalkanoate (PHA) as well as a construction method and application of the recombinant strain. The recombinant strain disclosed by the invention can be used for efficiently producing high-proportion poly (3-hydroxybutyrate-4-hydroxybutyric acid) (P34HB for short) by using glucose as a unique carbon source or a 4HB structure related or non-related carbon source in amplification production of a shake flask, a 7L fermentation tank, a 100L fermentation tank and a 5000L fermentation tank.

The invention relates to the technical field of biology, in particular to a beta-1, 4-galactosyl transferase mutant and a method for biologically synthesizing lactose-N-neotetraose by using the beta-1, 4-galactosyl transferase mutant. The invention discloses a protein, which is a protein HpgalT-186 or a protein HpgalT-146-186, a protein HpgalT-146 and a protein HpgalT-146. The protein HpgalT-186 is a protein obtained by mutating an amino acid residue at the 186th site of a wild type HpgalT; the protein HpgalT-146-186 is a protein obtained by carrying out mutation on an amino acid residue at the 186 site and an amino acid residue at the 146 site of wild type HpgalT. On the basis, a series of escherichia coli is constructed and optimized, the yield of the LNnT synthesized through shake flask fermentation of the finally obtained strain reaches 1.73-4.1 g / L, the highest yield in a 5L fermentation tank reaches 20-45.2 g / L, and efficient synthesis of the LNnT is achieved.

The invention includes a mutant Taq polymerase, which can significantly extend and amplify a target sequence where the extension conditions are time limited to as little as one second. The mutant Taq polymerase, or a biologically active fragment thereof, has one or more substitutions differing from the wild type as shown in Table I.

The invention belongs to the field of molecular biology, and particularly relates to a transgenic soybean event LP207-1 and a detection method thereof. The invention provides a nucleic acid sequence for detecting a transgenic soybean event LP207-1, the nucleic acid sequence is selected from one or more of sequences SEQ ID NO: 1-7 or complementary sequences thereof, and the nucleic acid sequence is derived from a plant, a seed or a cell of the transgenic soybean event LP207-1. A representative sample of a seed of the transgenic soybean event LP207-1 is preserved in the China Center for Type Culture Collection on May 23, 2025, and the preservation number is CCTCC NO: P202516. The transgenic soybean event LP207-1 has good resistance to lepidoptera pests and can tolerate agricultural herbicides containing glyphosate and glufosinate-ammonium, economic loss caused by the lepidoptera pests can be avoided, and the yield is not reduced; and the breeding efficiency is enhanced, and the molecular marker can be used for tracking a breeding population and transgenic insertion fragments in offspring of the breeding population. The detection method provided by the invention can quickly, accurately and stably identify the existence of the plant material derived from the transgenic soybean event LP207-1.

The present invention relates to a glucosamine synthase and its application in achieving plant luminescence. The glucosamine synthase can synthesize milkvine, and then synthesize luciferin through other proteins to make the plant luminescent. The results obtained by the present invention significantly improve the luminescence intensity of the plant, and through a mobile phone or camera lens, visible photos of luminous plants can be obtained without long-term exposure; the luminous phenotype of the plant can be easily observed with the naked eye.

Provided are S-adenosylmethionine synthetase mutants and the use thereof, belonging to the technical field of biological enzyme engineering. Site-directed combinatorial mutations are performed on positions 189, 200, 234, 266 and 371 of S-adenosylmethionine synthetase MAT to obtain a series of mutants with improved enzyme activity, wherein the specific enzyme activity of I189V / V266H and V266H / N371R is increased by more than 10 times relative to the parent. The yields of S-adenosylmethionine synthesized by whole cells of recombinant bacteria expressing mutants I189V / V266H / N371R, V266H / Q234N / N371R and I189V / V266H / Q234N / N371R all reach 1550 mg / L or more. The provided enzyme mutants have broad application prospects in the production of S-adenosylmethionine and the construction of genetically engineered microorganisms for S-adenosylmethionine.

The invention discloses an application of a medicago sativa MsDGK5 gene and a protein coded by the medicago sativa MsDGK5 gene in salt stress resistance of plants, and belongs to the technical field of plant genetic engineering. The nucleotide sequence of the medicago sativa MsDGK5 gene provided by the invention is as shown in SEQ ID NO. 1. According to the application, MsDGK5 gene overexpressed arabidopsis thaliana and medicago sativa transformation plants are constructed, growth and development indexes and physiological and biochemical parameters of the plants under salt stress are systematically measured, and it is proved that overexpression of the MsDGK5 gene can significantly improve the salt tolerance of the transgenic plants. Based on the experimental results, the key regulation effect of the MsDGK5 gene in plant salt tolerance is defined for the first time, an important gene resource is provided for crop salt tolerance genetic improvement, and a new technical approach is also provided for coping with the increasingly aggravated soil salinization problem.

Aspects of the disclosure relate to complexes and other aspects relate to formulations (e.g., aqueous, lyophilized forms) comprising such complexes comprising an oligonucleotide (e.g., useful for targeting DMPK) covalently linked to an antibody (e.g., anti-TfR1 antibody).

The invention relates to the technical field of biological catalysis, and discloses an L379A mutant enzyme for preparing rebaudioside I. The enzyme is obtained by the following mutations generated by UGT76G1: leucine of the 379th amino acid sequence is mutated into alanine; the enzyme can be applied to RA in-vitro conversion preparation of RI with higher utilization value, the conversion rate is higher than 50%, and the enzyme activity is remarkably improved by 7 times compared with the original enzyme catalytic conversion enzyme activity. The UGT76G1 mutant disclosed by the invention has the advantages that (1) the blank is filled, and a special enzyme catalyst capable of realizing efficient in-vitro synthesis of rebaudioside I (RI) is provided for the first time; 2) efficiency jump: the catalytic conversion rate is greatly increased from original about 7% to more than 50% (increase gt; and 3) stability and reliability: in the cross-scale reaction of 10mL to 5L, the catalyst has stable catalytic performance, shows excellent industrial application potential, and is suitable for popularization and application.

The invention relates to a glycosyl transferase UGTBL1 mutant with improved selectivity and application of the glycosyl transferase UGTBL1 mutant in catalytic synthesis of salidroside, and belongs to the field of gene engineering and enzyme engineering. A single-point mutation technology is adopted, the obtained mutant comprises 132-site amino mutation of an amino acid sequence, and the amino acid sequence of the glycosyltransferase is shown as SEQ ID NO: 1. Research results show that the glycosyl transferase mutants S132V and S132G have good selectivity, tyrosol and uridine diphosphate glucose can be efficiently catalyzed to synthesize the salidroside, the selectivity is 89%, and the glycosyl transferase mutants S132V and S132G are suitable for enzymatic preparation of the salidroside and have wide application prospects.

Aspects of the disclosure relate to complexes and other aspects relate to formulations (e.g., aqueous, lyophilized forms) comprising such complexes comprising an oligonucleotide (e.g., useful for targeting DMPK) covalently linked to an antibody (e.g., anti-TfR1 antibody).

There is provided a fluorinase variant thereof having at least 70% sequence identity to a sequence MSDLGX6TDDSVAQCKGLMLSICPX24VX26IX28DX30CHX33MTPX37DVVEGARYIVDLPR X52FPEGTVFATTTYPATGTX70X71RSVAX76RX78KX80AALGGARGQX90AGSGX95GX97E RAEGX103YIYIAPNNGLLTX116VIEEHGYX124EAYEVSX131TX133VIPX137X138PEPTFYSR EMVAIPSAHLAAGFPLX163X164VGRX168LX170DX172EIVRFEX179X180KX182X183X184VX186G X188X189LX191GX193X194X195X196X197DHPFGNX204WTNX208HRTDLEKAGIX219YX221TX223X 224KX226VX228DGVLX233FX235LPLX239PTFADAX246X247X248GX250PVX253YX255NSRGYLX2 62X263ARNAAX269LAYPYNLX277AGX280SVX283VTX286A (SEQ ID NO: 1), wherein X is a natural amino acid, and wherein the variant comprises one or more mutations at a site selected from the group consisting of an ion-egress site, a S-Adenosyl-L-Methionine (SAM) binding site, an ion-binding site (IBS), and a conserved site. Also disclosed are polynucleotides encoding the variants, vectors comprising the polynucleotides encoding the variants, and host cells comprising the vectors thereof. Also disclosed are Methods for producing the variants, methods of catalyzing the fluorination of a compound, uses of the fluorinase variants, and methods of treatment using the fluorinase variants thereof.

The present application relates to a method of producing a protein comprising an unnatural amino acid (UAA), the method comprising culturing a host cell, where the host cell is a eukaryotic cell, together with a nucleotide sequence encoding a first recoding tRNA or a first recoding tRNA, where the first recoding tRNA comprises an anticodon complementarily paired with a first codon, wherein the first codon is a rare codon; a nucleotide sequence encoding a first aminoacyl-tRNA synthetase or a first aminoacyl-tRNA synthetase, the first aminoacyl-tRNA synthetase being capable of loading a first recoding tRNA with a first non-natural amino acid.

The invention specifically discloses a composition for detecting NAD < + > and application thereof, and relates to the technical field of biological detection. According to the composition for detecting NAD < + >, the composition comprises an NAD < + > binding protein fragment A connected with an Fc tag, an NAD < + > binding protein fragment B and a connecting peptide, the detection result is accurate and reliable, and meanwhile the technical difficulty, uncertainty and high cost faced by antibody screening in a traditional method are avoided.