57915results about "Vector-based foreign material introduction" patented technology

Aspects of the disclosure relate to complexes comprising a muscle-targeting agent covalently linked to a molecular payload. In some embodiments, the muscle-targeting agent specifically binds to an internalizing cell surface receptor on muscle cells. In some embodiments, the molecular payload inhibits expression or activity of DUX4. In some embodiments, the molecular payload is an oligonucleotide, such as an antisense oligonucleotide or RNAi oligonucleotide.

A recombinant humanized collagen and a preparation method therefor. The recombinant humanized type III collagen can be efficiently and stably expressed in three expression systems of yeast, Escherichia coli, and CHO cells, and has the biological activity superior to that of a commercially available collagen product. The recombinant humanized type III collagen has structural characteristics and functional activity similar to those of a natural collagen, and thus has wide application prospects in biomedical materials, tissue engineering products, cosmetics, foods, health care products or drugs, medical instruments, medical cosmetology and the like.

The present invention discloses a human serum albumin-targeting nanobody and its application. A variety of human serum albumin-targeting nanobodies that can be used to construct long-acting fusion protein drugs and their immune screening methods are disclosed. The antibody is composed only of the heavy chain variable region and can be fused and expressed with therapeutic proteins or polypeptide drugs such as interleukin, interferon, and tumor necrosis factor in an Escherichia coli expression system. The fusion protein has the activity of specifically binding to human serum albumin, has a relatively high affinity, can effectively extend the drug half-life, and does not affect the biological activity of the recombinant protein drug at the same time.

The present application relates to a kind of deletion mutant nucleic acid and its application in anti-herbicide.The deletion mutation occurs in the promoter of rice OsHPPD Gene, the length of deletion sequence is at least 10 bp, and the A in the start codon ATG of gene is 0, at least the bases located in the upstream of start codon ATG from 2041 to 2032 are deleted. OsHPPD The present application finds that by deleting part of the sequence in the promoter of rice OsHPPD Gene, rice can obtain resistance to HPPD inhibitor herbicides, which is of great application value for ensuring agricultural production safety and improving the efficiency of herbicide use.

The invention belongs to the technical field of gene engineering, and particularly relates to a beta-glycosidase SS-BGL mutant for preparing rare ginsenosides and application of the beta-glycosidase SS-BGL mutant. The beta-glycosidase SS-BGL mutant is obtained by conducting single-site mutation or double-site mutation on beta-glycosidase SS-BGL with the amino acid sequence shown as SEQ ID NO.1. The beta-glycosidase SS-BGL mutant is obtained by conducting single-site mutation or double-site mutation on beta-glycosidase SS-BGL with the amino acid sequence shown as SEQ ID NO.2. Wherein the single site mutation is that phenylalanine at the 229 site of SEQ ID NO. 1 is mutated into proline; the double-site mutation is as follows: phenylalanine at the 229 site in SEQ ID NO. 1 is mutated into proline, and glycine at the 301 site is mutated into proline; or the phenylalanine at the 229 site in SEQ ID NO. 1 is mutated into proline, and the leucine at the 337 site is mutated into valine; or the phenylalanine at the 229 site in SEQ ID NO. 1 is mutated into the proline, and the glycine at the 338 site is mutated into the proline. On the basis of natural beta-glycosidase SS-BGL, the molecular structure of the beta-glycosidase SS-BGL is modified through site-directed mutagenesis, and four beta-glycosidase SS-BGL mutants with the saponin conversion activity improved are obtained.

The invention belongs to the technical field of cosmetics, and discloses anti-aging recombinant collagen as well as a preparation method and application thereof. The amino acid sequence of the anti-aging recombinant collagen is as shown in SEQ ID NO.1, and the nucleotide sequence of the anti-aging recombinant collagen is as shown in SEQ ID NO.2. The recombinant collagen CaI1 is constructed and expressed by recombining the effective sequences of the I-type collagen, the III-type collagen and the XVII-type collagen, and the recombinant collagen CaI1 is free of cytotoxicity, free of hidden danger of endotoxin and good in safety. The skin care product added with the recombinant collagen CaI1 can promote skin collagen absorption, inhibit skin aging and improve skin elasticity, and has a good application prospect.

The invention discloses a bovine serum albumin combined nano antibody and a preparation method and application thereof, the amino acid sequence of the bovine serum albumin combined nano antibody Nb1 is shown as SEQ.ID No.1, the nano antibody Nb not only has high affinity to bovine serum albumin, but also has good binding activity to both human serum albumin and mouse serum albumin, and the nano antibody Nb has high affinity to human serum albumin and mouse serum albumin. And the antibody has a relatively good broad-spectrum application range, and can be widely applied to the aspects of drug half-life period enhancement, tumor targeted therapy and imaging, immunodetection or diagnosis, recombinant albumin purification or enrichment and the like.

Aspects of the disclosure relate to methods of reducing fatigue in a subject having myotonic dystrophy type 1 (DM1). Aspects of the disclosure relate to methods of treating one or more symptoms assessable by the MDHI (e.g., a GI symptom, myotonia, upper extremity function impairment, fatigue, mobility impairment, impairment in the ability to perform activities, pain, vision impairment, communication impairment, sleep impairment, emotional issues, cognitive impairment, social satisfaction impairment, social performance impairment, breathing impairment, swallowing impairment, and / or hearing impairment) in a subject having myotonic dystrophy type 1 (DM1). 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-TfRl antibody).

This invention provides a halogenated compound for 5' end capping of nucleic acids and its applications, the compound being shown in formula (I). This invention also provides the uses and effects of the above-described compound on nucleic acid transcription and expression.

The invention belongs to the technical field of biochemical engineering, and provides a method for increasing L-carnosine yield of escherichia coli by increasing PRPP supply, and the yield and yield of L-carnosine are remarkably increased by up-regulating expression of a prs gene, increasing PRPP content in cells and promoting synthesis of L-histidine. Experiments show that the yield of the L-carnosine in different recombinant strains is increased by 55.56%-140%, and L-histidine and beta-alanine do not need to be added in the production process.

The invention relates to the technical field of biology, in particular to a verticillium dahliae virulence gene, a verticillium dahliae virulence protein and application. The invention discloses a verticillium dahliae virulence gene VdPHO23like. The verticillium dahliae virulence gene VdPHO23like comprises a polynucleotide sequence for coding an amino acid sequence of SEQ ID NO: 3. According to the invention, expression of the gene is inhibited in a targeted manner through an RNA interference mediated gene silencing technology so as to weaken virulence of pathogenic bacteria, and an efficient, specific and environment-friendly comprehensive prevention and control strategy is provided for verticillium wilt of crops such as cotton.

The invention belongs to the technical field of gene engineering, and particularly relates to applications of OpgG and a mutant thereof in production of human milk oligosaccharide. The invention provides a dextran biosynthetic protein OpgG mutant, and the OpgG mutant is obtained by mutating the 443rd amino acid from threonine to proline on the basis of a wild type dextran biosynthetic protein OpgG as shown in SEQ ID NO.1. The invention also provides a preparation method of the dextran biosynthetic protein OpgG mutant. An opgG gene and / or a T443P mutant coding gene are / is introduced into a human milk oligosaccharide production strain and applied to production of human milk oligosaccharide, and the yield of 3 '-SL, 6'-SL, LNT II, LNnT and LNT is greatly increased.

The invention discloses an AsMYB19 gene and application thereof, and relates to the technical field of biology, the nucleotide sequence of the AsMYB19 gene is shown as SEQ ID NO.1, the AsMYB19 gene can regulate and control the tiller number and the yield of oat, gene resources and strategies are provided for oat breeding and improvement of oat germplasm resources, the breeding speed is increased, and the breeding efficiency is improved.

The present invention relates to the fields of genetic engineering and plant genetic breeding, and specifically to the application of wheat TaPND gene. In the present invention, by taking gene TaPND as a target, designing a sgRNA sequence based on CRISPR / Cas9, connecting a DNA fragment containing the encoding sgRNA sequence to a vector carrying CRISPR / Cas, a plant binary expression vector capable of knocking out the TaPND gene in wheat is constructed; by genetically transforming wheat, offspring with TaPND gene knockout are obtained, and wheat mutants with early flowering time are obtained. Experiments have confirmed that transgenic wheat with TaPND gene knockout achieves the advancement of wheat flowering time. The technical scheme of the present invention provides a practical method for realizing rapid wheat breeding using genetic engineering technology, which has important breeding application value and broad market application prospects.

The invention provides a carbonyl reductase mutant. The carbonyl reductase mutant is obtained by performing single-point mutation or multi-point combined mutation on the 17th site, the 40th site and the 64th site of an amino acid sequence as shown in SEQ ID NO.1. The invention also provides a coding gene, a recombinant vector containing the coding gene, a co-expression engineering bacterium and an application of the co-expression engineering bacterium. Compared with a wild type enzyme, the carbonyl reductase mutant has high activity and high stereoselectivity, the enzyme activity can reach more than two times that of the wild type enzyme, the catalytic efficiency on a substrate precursor ketone is remarkably improved, the yield of a statin drug intermediate and chiral alcohol synthesized by an enzyme method of the carbonyl reductase mutant is remarkably improved, and the yield of a rosuvastatin intermediate (3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R) is remarkably improved. The highest yields of the (3R, 5R)-6-chloro-3, 5-dihydroxyhexanoic acid tert-butyl ester and the atorvastatin intermediate (3R, 5R)-6-cyano-3, 5-dihydroxyhexanoic acid tert-butyl ester reach 99% and 97.6% respectively, e.e. Is larger than 99%, and the method has high industrial application value.

The invention provides humanized recombinant XVII collagen for promoting hair follicle stem cell repair and resisting aging as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. The humanized recombinant XVII type collagen comprises polypeptide derived from the XVII type collagen or repeated protein formed by N times of repeated series connection of the polypeptide serving as a unit, n is an integer from 2 to 4; the polypeptide derived from the XVII type collagen comprises a polypeptide A and / or a polypeptide B; the amino acid sequence of the A polypeptide is as shown in SEQ ID NO: 1; and the amino acid sequence of the B polypeptide is as shown in SEQ ID NO: 2. The humanized recombinant XVII type collagen not only can be massively and stably recombined and expressed, but also has cell adhesion activity, cell proliferation promotion activity, cell migration promotion activity and excellent hair follicle repair and regeneration promotion activity, and has no sensitization side effect. The invention provides a new way for preparing collagen products with rich functions.

The invention relates to a recombinant A type humanized XVII type collagen as well as construction and application thereof, the protein is the humanized XVII type collagen which is composed of an intracellular region, a transmembrane region and an extracellular region, and the amino acid sequence is as shown in SEQ No. 1. The hair follicle repairing agent has the advantages of hair follicle repairing, hair regeneration, good biological activities of cell proliferation, cell adhesion and the like, and can be widely applied to the fields of medicines, medical instruments, biological materials, tissue engineering, cosmetics and the like. The gene sequence of the recombinant A type XVII collagen is 100% identical to the corresponding part of the gene sequence of the recombinant A type XVII collagen, so that the recombinant A type XVII collagen has good hydrophilicity and high activity, and has application potential in the fields of biomedical materials, beauty cosmetics, skin care products and the like.

Described herein are systems and methods for automated cell transduction within a cell processing system. A system for cell processing may include a cell processing cartridge having a transduction system. The transduction system may include a fluidic manifold, one or more modules for performing a cell processing protocol, and a tube having a surface area to volume ration of between about 1,260 mm2 / mL and about 5,080 mm2 / mL. A method for cell processing may first include flowing cells through a tube of a flow cell of a cell processing cartridge for a first time period to achieve a transduction efficiency of at least 50%. Second, the method may include expanding the cells within a bioreactor module of the cell processing cartridge for a second time period.

The invention relates to the field of rice gene engineering, and particularly provides OsSULTR2; the invention also discloses application of the 2 protein and the coding gene thereof in regulating and controlling the salt tolerance of rice. The protein meets the following conditions: B1) a protein with an amino acid sequence of SEQ ID NO.1; and B2) a fusion protein with the same function obtained by connecting a tag to the N end and / or C end of B1). The method is used for detecting OsSULTR2; the salt stress phenotype identification in the seedling stage is carried out on the transgenic rice with the gene knockout 1, 2, and the result shows that when the gene segment is deleted, the salt stress tolerance of the rice is improved, and the function and the application way of the gene are proved. Therefore, the OsSULTR2 of the present invention; the 2 protein and the coding gene thereof can regulate and control the salt tolerance of rice, and have important significance for cultivating salt-tolerant transgenic rice.

The invention provides compositions and methods for treating diseases such as cancer. The invention also relates to a method of administering a therapy comprising a chimeric antigen receptor, an IL-15R molecule and an IL-15 molecule.

The invention discloses a high temperature resistant xanthan gum lyase mutant and a preparation method thereof, wild type xanthan gum lyase comes from Paenibacillus anensis, site-directed mutagenesis is carried out on the wild type xanthan gum lyase to obtain the high temperature resistant xanthan gum lyase mutant, and the mutant comprises at least one of N66V, R292A and S676V. The enzymatic activity of the xanthan gum lyase mutant treated in a buffer solution with the pH value of 5.0 at the temperature of 65-85 DEG C for 2 h is remarkably improved compared with that of an original enzyme, and the enzymatic activity of N66V / R292A / S676V is improved by 3.8 times compared with that of the original enzyme at the temperature of 85 DEG C.

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 relates to the field of gene engineering, and discloses a D-3-phosphoglycerate dehydrogenase mutant, a coding gene thereof and application of the D-3-phosphoglycerate dehydrogenase mutant and the coding gene. Compared with a protein with an amino acid sequence as shown in SEQ ID NO: 1, the mutant comprises mutation at the 131 site and / or the 285 site. The mutant can specifically catalyze a substrate 3-phosphoglycerate to synthesize 3-phosphate-hydroxypyruvic acid, can effectively promote synthesis of L-serine in a metabolic pathway of microorganisms, improves the yield and conversion rate of the L-serine, and reduces the yield of byproducts at the same time.

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 invention provides a modified TnpB-omega RNA gene editing system and application of the modified TnpB-omega RNA gene editing system. In particular, the present invention provides a modified TnpB related [omega] RNA skeleton. The modified omega RNA skeleton provided by the invention has at least three stem-loop structure fragments, and has a shorter length and a compact structure than a natural omega RNA skeleton, so that the modified omega RNA skeleton and TnpB can be jointly carried in a single AAV vector, the administration efficiency is improved, and high-efficiency gene editing is realized.

The invention provides an artificial nucleic acid molecule which is used for improving the expression quantity of target amino acid, polypeptide or protein. The artificial nucleic acid molecule at least comprises a target 5'untranslated region (UTR), a target coding region (CDS) and a target 3 'untranslated region (UTR). Wherein the sequence of the target 5 'UTR is one of the following sequences: 5' UTR of a high-expression gene and a 5 'UTR variant of the high-expression gene. The sequence of the target 3 'UTR is one of the following sequences: 3' UTR of a high-expression gene and a 3 'UTR variant of the high-expression gene. Optionally, the artificial nucleic acid molecule may further comprise, for example, a 5 '-end cap structure (Cap), a PolyA tail. The 5 'UTR and the 3' UTR have regulating effects on translation and stability of nucleic acid molecules, so that the 5 'UTR, the 3' UTR and variants thereof are selected from high-expression genes, the nucleic acid molecules can be further stabilized and are not easy to degrade, and the amount of protein or polypeptide obtained by translation of the nucleic acid molecules can be increased. The invention also provides methods for making, delivering, and using such artificial nucleic acid molecules, as well as the use of the artificial nucleic acid molecules for the treatment and / or prevention of related diseases or disorders.

The invention discloses a site for stably expressing protein in a CHO cell gene NW023276806.1 and application of the site, and belongs to the technical field of biological genes. The site belongs to a fixed position in a CHO cell genome, different protein genes are introduced based on a micro-homologous end connection mechanism through a CRISPR / Cas9 tool, and stable expression is carried out. By adopting a site-specific integration method, a target gene is integrated to a stable expression area in a site-specific manner, repeated high-expression monoclonal screening is effectively avoided, and an MMEJ mechanism is introduced to integrate a donor fragment, so that the research and development time for constructing a stable expression cell strain in biological pharmacy can be effectively shortened, and the cost is reduced.