75606results about "Vector-based foreign material introduction" patented technology

The embodiments of the present disclosure relate to decreasing the bioavailability of one or more target biomolecules by providing a composition that comprises a recombinant plasmid with one or more sequences of micro interfering ribonucleic acid (miRNA). When the recombinant plasmid interacts with a target cell, it causes the target cell to upregulate production of the miRNA, which then decreases the bioavailability of the target biomolecule. In some embodiments of the present disclosure, the target biomolecule is a cytokine or other mediator molecule of an inflammatory response.

The embodiments of the present disclosure relate to one or more compositions or methods that upregulate the production of one or more sequences of micro-interfering ribonucleic acid (miRNA). The sequences of miRNA may be complimentary to a sequence of target messenger RNA (mRNA) that encodes for translation of a target biomolecule, such as JAK. The miRNA can cause the target mRNA to be degraded or inactivated, thereby causing a decrease in bioavailability of the target biomolecule because it is degraded or inactivated by the miRNA. Decreasing the bioavailability of the target biomolecule within a subject that is administered the one or more compositions may address the afflictions experienced by the subject due to expression of the target biomolecule.

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.

The present application relates to the field of biotechnology, and in particular to an epimerase for synthesizing D-tagatose and its application, wherein the epimerase produces a mutation at position 100, or at position 125, or at position 127, or at position 131, or at position 268, or at position 270, or at position 271, or at position 308, or at position 339, or at position 340, or at position 342, or at position 366, or at position 402, or at position 437, or at position 440 in the amino acid sequence shown in SEQ ID NO. 142; through the above-mentioned method, the epimerase exhibits excellent epimerization activity at the 4th carbon position of D-fructose, can efficiently convert D-fructose into D-tagatose, and is beneficial to improving the synthesis efficiency of D-tagatose.

The invention relates to the improvement of endonuclease-based antimicrobials by blocking DNA repair of double-strand break(s) (DSB(s)) in prokaryotic cells. In this respect, the invention especially concerns a method involving blocking DNA repair after a nucleic acid has been submitted to DSB, in particular by a Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) associated programmable double-strand endonuclease. The invention particularly relates to the use of an exogenous molecule that inhibits DNA repair, preferably a protein that binds to the ends of the double-stranded break to block DSB repair. The invention also relates to vectors, particularly phagemids and plasmids, comprising nucleic acids encoding nucleases and Gam proteins, and a pharmaceutical composition and a product containing these vectors and their application.

The invention provides a preparation method of a novel recombinant humanized collagen hydrogel. The method comprises the following steps: recombinant humanized collagen and a dialdehyde crosslinking agent are subjected to a reaction, the concentration of the dialdehyde crosslinking agent is 0.01-0.25%, and the recombinant humanized collagen is one or more of recombinant humanized type II collagen, recombinant humanized type III collagen and recombinant humanized type XVII collagen, and the recombinant humanized collagen has a triple helix structure. The aldehydes cross-linking agent suitable for the recombinant humanized collagen, especially the recombinant humanized collagen with a triple helix structure, and the optimal reaction conditions are determined.

The invention belongs to the technical field of gene engineering and enzyme engineering, and particularly relates to an ornithine decarboxylase mutant as well as a coding gene, an expression vector, a recombinant strain and application thereof. The ornithine decarboxylase mutant is obtained by mutating wild ornithine decarboxylase ODC10 of which the amino acid sequence is shown as SEQ ID NO.1, the catalytic activity of the obtained mutant is improved, and the biosynthesis efficiency of 1, 4-butanediamine is favorably improved. The recombinant strain for expressing the ornithine decarboxylase mutant constructed by the invention can efficiently synthesize 1, 4-butanediamine, and the molar conversion rate of ornithine is remarkably improved, so that the production benefit is improved, and the production cost is reduced.

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.

To provide VHH-containing polypeptides that bind CD123, and a method for treating cancer that includes administration of the VHH-containing polypeptides.SOLUTION: Provided is a polypeptide comprising at least one VHH domain that binds to CD123, comprising CDR1, CDR2, and CDR3, each comprising a specific amino acid sequence.SELECTED DRAWING: Figure 3A

The invention belongs to the technical field of bioengineering, and discloses a recombinant yarrowia lipolytica capable of efficiently and rapidly producing sclareol, which is characterized in that optimized sclareol synthase related coding genes ScLPPS, ScSCS, GGPPS1, GGPPS2, ERG20 and tHMGR coding genes are inserted into a yarrowia lipolytica genome; inserting optimized pentadienol utilization pathway coding genes such as IPK, THIM and IDI coding genes; and inserting an optimized ATP: citrate lyase coding gene ACLY to obtain the ATP: citrate lyase coding gene ACLY. The recombinant yarrowia lipolytica is based on knockout of a yarrowia lipolytica strain responsible for coding a non-homologous recombination gene ku70, so that the homologous recombination capability of the yarrowia lipolytica strain is enhanced, gene integration is realized through the homologous recombination function of the yarrowia lipolytica, the genetic stability of the introduced gene can be greatly improved, and the recombinant yarrowia lipolytica strain has a good application prospect. The construction method of the recombinant yarrowia lipolytica strain is efficient and simple to operate.

Methods for making synthetic gene clusters are described.

The invention relates to a recombinant escherichia coli THRS and a method for synthesizing L-threonine by using the recombinant escherichia coli THRS. The invention belongs to the technical field of bioengineering, and provides a recombinant escherichia coli THRS which overexpresses encoding genes of ppc, aspC, aspA, thrAmutBC, asd, rhtC, rhtA, asdtm and thrAmut, knockout an encoding gene of fnr and knockout an encoding gene of iclR, the recombinant escherichia coli THRS takes glucose as a substrate for fermentation, and the yield and conversion rate of L-threonine are obviously improved; the L-threonine strain has the advantages that the L-threonine strain does not contain exogenous plasmids, all modification sites are integrated onto a genome, higher stability is achieved, antibiotics do not need to be added into a fermentation culture medium of the strain, and the produced L-threonine product is free of antibiotic residues and higher in safety.

To provide methods for treating a subject having or at risk of developing a neurocognitive disorder, such as frontotemporal lobar degeneration or neuronal ceroid lipofuscinosis.SOLUTION: Provided is a method comprising administering, to a subject diagnosed as having a neurocognitive disorder (NCD), a composition comprising a population of cells comprising a transgene encoding a progranulin (PGRN) or a granulin (GRN).SELECTED DRAWING: Figure 1A

To provide a medicine for treating a disease in a mammal.SOLUTION: Provided is a medicine for treating a disease in a mammal, comprising an expression cassette that is a first expression cassette comprising a chimeric gene operably linked to a first expression control element, the chimeric gene comprising a first portion comprising a minigene having an alternatively spliced exon and a second portion encoding a first RNA encoding a protein, the first portion being 5' to the second portion, the second portion lacking an initiation codon or start codon, the alternatively spliced exon comprising a translation initiation regulatory sequence, expression of the protein being controlled by inclusion of the alternatively spliced exon, and the medicine being used in combination with a small molecule splicing modifier that modulates splicing of the alternatively spliced exon.SELECTED DRAWING: Figure 1

To provide constructs, vectors, relative host cells and pharmaceutical compositions which allow an effective gene therapy, in particular of genes larger than 5Kb.SOLUTION: The present invention implements cellular large protein reconstitution by providing to a target cell two or more fragments of large protein fused to split inteins to promote intein-mediated trans-splicing and reconstitute the functional protein.SELECTED DRAWING: None

The invention provides a multielement polyA polynucleotide for improving mRNA (messenger ribonucleic acid) stability and protein expression and application of the multielement polyA polynucleotide. The invention firstly provides a polynucleotide, the polynucleotide comprises more than two sections of polyA sequence fragments, the adjacent polyA sequence fragments are connected through Linkers, each polyA sequence fragment is composed of one or more continuous A, and each Linker is composed of C and / or G. The polynucleotide provided by the invention can improve mRNA stability and protein expression, and compared with a traditional unit poly-A sequence, the polynucleotide provided by the invention can greatly reduce the recombination rate of fermentation in escherichia coli so as to more easily optimize and amplify the process, and can be better applied to plasmid fermentation related to transcription template preparation in mRNA vaccine or drug production.

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.

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.

The invention relates to recombinant escherichia coli THRS-6 and a method for synthesizing L-threonine by using the recombinant escherichia coli THRS-6. The invention belongs to the technical field of bioengineering, and relates to an aspartic acid kinase thrAmut and pyridine nucleotide transhydrogenase pntAB gene derived from escherichia coli, an aspartic acid dehydrogenase aspDH gene derived from pseudomonas aeruginosa, a hemoglobin vgb gene derived from vitreoscilla and a transcriptional regulation factor irrE gene derived from radiococcus. The recombinant escherichia coli is used as a substrate, the ldhA gene is knocked out to further modify the recombinant escherichia coli, the recombinant escherichia coli THRS-6 with glucose as the substrate is obtained, the THRS-6 is used for fermentation, and the yield and conversion rate of the L-threonine are obviously improved.

The invention belongs to the technical field of microorganisms, and particularly relates to recombinant escherichia coli containing a PDRN sequence and a preparation method thereof, and the preparation method comprises the following steps: S1, preparing a PDRN solution; s2, complementing the double strands of the PDRN into blunt ends; s3, constructing a recombinant plasmid; s4, carrying out mixed culture on a product obtained in the step S3 and escherichia coli DH5alpha; s5, screening the recombinants; s6, carrying out PCR (Polymerase Chain Reaction) amplification, and recording a strain from a fragment with the length of more than 200bp; s7, preparing a seed solution by adopting the strain prepared in the step S6; s8, strain fermentation; and S9, centrifugally collecting thalli. The method comprises the following steps: constructing a recombinant plasmid containing a PDRN fragment, introducing the recombinant plasmid into Escherichia coli, subsequently culturing the recombinant Escherichia coli to copy DNA in the Escherichia coli, extracting a purified exogenous gene fragment to obtain a strain containing a single fragment PDRN sequence, and getting rid of dependence on raw materials during subsequent extraction.

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 relates to a nano antibody targeting cadherin 17 and application thereof, and relates to the technical field of immunology and molecular biology, the amino acid sequence of a complementary determining region of the nano antibody targeting cadherin 17 comprises CDR1 shown as any one of SEQ ID NO: 21-32, CDR2 shown as any one of SEQ ID NO: 34-45, and CDR3 shown as any one of SEQ ID NO: 47-58. According to the invention, an alpaca immune phage display library is constructed, a series of nano antibody molecules targeting cadherin 17 (CDH17) are screened from the alpaca immune phage display library, and functions and applications of the nano antibody are verified at various levels of CDH17 antigen protein, CDH17 target cells, CDH17 CAR-T cells and the like.

The present invention belongs to the technical field of immunology, and in particular, relates to a humanized nanoantibody against TNF-α and its application. The present invention provides a humanized nanoantibody against TNF-α, and a recombinant humanized nanoantibody against TNF-α prepared by expressing the gene of the humanized nanoantibody against TNF-α. The humanized nanoantibody against TNF-α prepared by the present invention can specifically recognize the TNF-α antigen, thereby accurately targeting the inflammatory site, reducing nonspecific reactions, and improving the therapeutic effect.

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 present disclosure provides guided microbial remodeling (GMR) methods for the rational improvement of plant-associated microbes to perform plant-beneficial functions. The GMR methods described herein allow for non-intergeneric genetic optimization of key regulatory networks within the microbes, which improve plant-beneficial functions over wild-type microbes but don't have the risks associated with transgenic approaches (e.g., unpredictable gene function, public and regulatory concerns, etc.). The present disclosure also provides remodeled microbes and compositions thereof. The utilization of remodeled microbes and compositions thereof will enable farmers to realize more productive and predictable crop yields without the nutrient degradation, leaching, or toxic runoff associated with traditional synthetically derived fertilizers.

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 belongs to the technical application field of microbial metabolic engineering and synthetic biology, and particularly relates to a pichia pastoris engineering strain for high-yield sclareol as well as a construction method and application of the pichia pastoris engineering strain. The method comprises the following steps: constructing a biosynthetic pathway of sclareol in a host strain, and optimizing an intracellular mevalonic acid metabolic pathway and a central metabolic pathway to obtain an engineering strain A; the host strain is pichia pastoris; or overexpressing or knocking out a metabolism regulation factor in the obtained engineering bacterium A to obtain an engineering bacterium strain B; or, in the obtained engineering bacterium B, a synthetic pathway is targeted to peroxisome and optimized by adopting a cell region compartment strategy, and an engineering bacterium strain C is obtained. The invention provides a pichia pastoris chassis cell for synthesizing a diterpenoid sclareol and an engineering strain for synthesizing sclareol. By introducing and optimizing the expression of a sclareol synthesis pathway, the fermentation yields of sclareol in shake flask batch and bioreactor batch fed-batch fermentation are respectively 631.6 mg / L and 10.5 g / L.