6549results about "Nucleic acid vector" patented technology

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 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.

The invention belongs to the technical field of biological medicines, and provides application of DRAM1 as a diagnosis and treatment target for resisting non-alcoholic fatty liver disease related hepatocellular carcinoma in preparation of a kit, a carrier and a medicine aiming at the problem of lack of effective treatment targets of NAFLD-related HCC. As a target spot for resisting the non-alcoholic fatty liver disease related hepatocellular carcinoma, the expression level of the DRAM1 in liver tissues of non-alcoholic fatty liver disease related hepatocellular carcinoma patients and mice is increased. According to the application disclosed by the invention, research is developed aiming at the effect of the DRAM1 in the generation and development of NAFLD-related HCC, the analysis of TCGA liver cancer queue data and a GSE164760 data set is taken as an entry point, the influence of the DRAM1 on the diagnosis and prognosis of the liver cancer is comprehensively analyzed, and verification is carried out through a mouse in-vivo experiment, so that a new thought and a new target spot are provided for clinical diagnosis and treatment of the liver cancer.

The specification provides methods for introducing a desired genetic change in a nucleotide sequence using a double-strand break (DSB)-inducing genome editing system, the method comprising: identifying one or more available cut sites in a nucleotide sequence; analyzing the nucleotide sequence and available cut sites with a computational model to identify the optimal cut site for introducing the desired genetic change into the nucleotide sequence; and contacting the nucleotide sequence with a DSB-inducing genome editing system, thereby introducing the desired genetic change in the nucleotide sequence at the cut site.

The invention provides a recombinant XVII type collagen and application thereof, and relates to the technical field of protein engineering. The novel recombinant human XVII type collagen is obtained by optimizing and screening collagen structural domain sequences of the human XVII type collagen and then combining the sequences. The recombinant human XVII type collagen is consistent with the corresponding part of the amino acid sequence of the human XVII type collagen, does not generate immunological rejection when being applied to a human body, and can be widely applied to the industries of surgical medical treatment and medical beauty. In addition, the recombinant human XVII type collagen is high in stability, and the problem of protein degradation in the process of expressing recombinant protein by pichia pastoris can be avoided. In addition, the recombinant human XVII type collagen also has the effects of high biological activity, high biological safety, promotion of cell proliferation and differentiation and promotion of hair growth, and can be applied to the field of cosmetics or medicines as a functional component.

The embodiment of the invention provides a method for obtaining a tyrosine ammonialyase mutant based on high-throughput screening, the tyrosine ammonialyase mutant and application of the tyrosine ammonialyase mutant. The mutant is subjected to any one of the following amino acid mutations on an amino acid sequence as shown in SEQ ID NO.2: A245G + A247T, and D192V + A245G + A247T. According to the embodiment of the invention, an amino acid sequence as shown in SEQ ID NO.2 is taken as a template (basis), a key active site of the template is subjected to mutation transformation, a mutant library is constructed, and a series of mutants with remarkably improved catalytic activity and / or conversion rate are screened; the tyrosine ammonia lyase mutant provided by the embodiment of the invention can selectively catalyze L-tyrosine to generate p-coumaric acid, and has excellent yield and / or higher yield, which indicates that the mutant has more significant application prospects in catalysis of L-tyrosine to generate p-coumaric acid.

The present disclosure provides an antibody or antigen-binding fragment thereof that specifically binds to human cMET. The disclosure also provides polynucleotides encoding the antibodies or antigen binding fragments thereof, vectors comprising the polynucleotides, host cells comprising the vectors, methods of generating the antibodies, and compositions comprising the antibodies.

The invention provides a UTR (Untranslated Region) sequence for improving mRNA (Messenger Ribonucleic Acid) stability and translation efficiency and application of the UTR sequence, and particularly provides an mRNA molecule which comprises a coding region for coding polypeptide or protein and a fragment thereof, a 5 'UTR sequence positioned at the upstream of the coding region and / or a 3' UTR sequence positioned at the downstream of the coding region, the 5 'UTR sequence is a nucleotide sequence as shown in SEQ ID NO: 1, and the 3' UTR sequence is a nucleotide sequence as shown in SEQ ID NO: 2; and / or, the 5 'UTR sequence is a nucleotide sequence as shown in SEQ ID NO: 3, and the 3' UTR sequence is a nucleotide sequence as shown in SEQ ID NO: 4. According to the present invention, the mRNA stability and the translation efficiency are improved through the new and optimized UTR sequence, and the UTR sequence can be used as the element for enhancing the RNA expression efficiency in the nucleic acid treatment drug or the mRNA vaccine so as to provide more and better choices for the mRNA therapy.

The invention discloses a multi-effect repair type collagen and application thereof, and belongs to the technical field of synthetic biology. Transdermal peptide TD-1, a transmembrane region of human XVII type collagen, an extracellular sixteenth non-collagen region, an extracellular fifteenth collagen region and a coding gene of a His tag are selected to be connected in series, and a gene sequence is optimized through pichia pastoris codon selection preference; and then the T-COL17R3 is constructed and expressed to obtain the T-COL17R3. Efficacy experiments prove that compared with similar products sold in the market, T-COL17R3 not only has excellent transdermal performance, but also has better cell proliferation and migration promoting capability, better capability of resisting early saccharification product ketoamine and later saccharification product dicarbonyl compounds, better capability of scavenging free radicals and resisting elastase, and can be used for preparing the anti-epidermal drug. In addition, synthesis of melanin in B16-F10 cells can be reduced, so that T-COL17R3 has excellent application potential in development of pharmaceutical compositions or skin care products with repairing, anti-wrinkle and firming or whitening and brightening effects.

The invention provides a micromolecular recombinant XVII type collagen as well as a preparation method and application thereof, and relates to the technical field of protein engineering. The novel recombinant human XVII type collagen is obtained by optimizing and screening a plurality of spiral region sequences such as a collagen structural domain, a C-terminal region and a middle region of the human XVII type collagen. The amino acid composition of the recombinant XVII type collagen is consistent with that of a natural collagen alpha1 chain, immunological rejection is not generated when the recombinant XVII type collagen is applied to a human body, the recombinant XVII type collagen can be widely applied to surgical medical treatment and medical beauty industries, the molecular weight of the collagen is small, efficient secretion soluble expression in eukaryotic host cells such as pichia pastoris can be achieved, and the recombinant XVII type collagen has a good application prospect. And industrial large-scale production is carried out. According to the present invention, the verification results show that the micromolecular recombinant XVII type collagen further has basement membrane anti-aging activity, wherein the basement membrane anti-aging activity specifically comprises the improvement of the expression levels of the IV type collagen, the VII type collagen and the XVII type collagen;

Systems and methods for targeted gene modification, targeted insertion, perturbation of gene transcripts, and nucleic acid editing. Novel nucleic acid targeting systems comprise components of CRISPR systems and non-LTR retrotransposon elements.

Some aspects of this disclosure provide strategies, systems, reagents, methods, and kits that are useful for the targeted editing of nucleic acids, including editing a single site within the genome of a cell or subject, e.g., within the human genome. In some embodiments, fusion proteins of Cas9 and nucleic acid editing proteins or protein domains, e.g., deaminase domains, are provided. In some embodiments, methods for targeted nucleic acid editing are provided. In some embodiments, reagents and kits for the generation of targeted nucleic acid editing proteins, e.g., fusion proteins of Cas9 and nucleic acid editing proteins or domains, are provided.

The present disclosure provides compositions and methods of use thereof for targeting trans- splicing of a pre-mRNA in a cell utilizing small nuclear RNAs (snRNAs) and small nucleolar RNAs (snoRNAs), and related methods.

The invention relates to the technical field of microorganisms, and particularly discloses a recombinant microorganism and application thereof in fermentation production of decarboxylated carnosine. Compared with an original strain, the recombinant microorganism disclosed by the invention has the advantages that the expression of the mdtK gene is enhanced; the starting strain is escherichia coli capable of producing decarboxylated carnosine. When the recombinant microorganism is used for fermentation production, the yield of the decarboxylated carnosine can be increased, L-histidine, histamine or beta-alanine and other precursors do not need to be added in the production process, and an effective novel method is provided for industrial production of the decarboxylated carnosine.

The present invention is related to the field of protein engineering technology which increases the enzymatic activity and thermal stability of human hyaluronidase which is an enzyme that hydrolyzes hyaluronic acid; and more particularly to hyaluronidase PH20 variants or fragments thereof, which comprise one or more amino acid residue substitutions in the region corresponding to the alpha-helix region and its linker region in the amino acid sequence of wild-type PH20 of SEQ ID NO: 1 and in which one or more amino acid residues at the N-terminus and / or the C-terminus are selectively cleaved additionally.Specifically, the present invention relates to PH20 variants or fragments thereof, which comprise one or more amino acid residue substitutions selected from the group consisting of T341A, T341C, T341G, S343E, M345T, K349E, L353A, L354I, N356E and I361T in wild-type PH20 having the amino acid sequence of SEQ ID NO: 1, and additionally comprise the substitution of amino acids located in the alpha-helix 8 region and / or a linker region between alpha-helix 7 and alpha-helix 8 in the amino acid sequence of wild-type PH20, and in which one or more amino acids located at the N-terminal and C-terminal regions are deleted.

The invention discloses a high-transdermal recombinant humanized XVII type collagen as well as a preparation method and application thereof, and belongs to the technical field of synthetic biology. According to the preparation method, transdermal peptide TD-1, different functional fragments of human XVII type collagen and coding genes of His tags are selected to be connected in series, gene sequences are optimized through pichia pastoris codon selection preference firstly, and then the high-transdermal recombinant humanized XVII type collagen is obtained; and then the recombinant humanized XVII type collagen T-COL17-NC16 and the recombinant humanized XVII type collagen T-COL17 are constructed by expression of the recombinant humanized XVII type collagen T-COL17. Effect experiments prove that compared with similar products sold in the market, the skin-care product has more excellent transdermal performance, can improve cell proliferation and migration ability, also has obvious ability of resisting early saccharification product ketoamine and late saccharification product dicarbonyl compounds, better ability of scavenging free radicals and resisting elastase, and can be used for preparing the skin-care product. Moreover, synthesis of melanin in B16-F10 cells can be reduced, and the compound is expected to become a novel functional biological raw material to be applied to the fields of pharmaceutical compositions or skin care products and the like.

The invention belongs to the technical field of biology, and particularly relates to a mouse hepatitis virus antibody or an antigen binding fragment thereof as well as a preparation method and application thereof. The antibody or the antigen binding fragment thereof can specifically recognize and bind to the mouse hepatitis virus or the NSP3 protein thereof, and has good affinity with the mouse hepatitis virus or the NSP3 protein thereof; the kit can be used for preparing products for diagnosing, preventing and / or treating mouse hepatitis virus infection or diseases caused by the mouse hepatitis virus infection, detecting existence or level of mouse hepatitis virus or NSP3 protein in a sample, screening medicines for preventing and / or treating the mouse hepatitis virus infection or the diseases caused by the mouse hepatitis virus infection, and carrying out basic research and / or vaccine development.

The present invention provides a fucosyltransferase polypeptide, which can be applied to the synthesis of human milk oligosaccharides (HMOs) and belongs to the field of enzyme engineering technology. The fucosyltransferase polypeptide described in the present application comprises a polypeptide having at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99%, and 100% identical amino acid sequence to the polypeptide shown in SEQ ID NO:1-25. The transferase polypeptide described in the present application improves the ability of the polypeptide shown in SEQ ID NO:1 to catalyze the synthesis of 2'-FL while also improving the specificity of the polypeptide catalyzing the substrate.

The invention discloses a small CRISPR (clustered regularly interspaced short palindromic repeats)-Cas gene editing system and application thereof. According to the invention, based on microbial genome and metagenome data, a class of CRISPR-Cas family protein is mined through a biological information method, and is named as Cas12r. A CRISPR-Cas12r editing tool constructed on the basis of the gene can realize gene editing in prokaryotic or eukaryotic cells. The CRISPR-Cas12r gene editing system obtained by the invention has the characteristics of miniaturization and various PAM types.

Strategies, systems, methods, reagents, and kits for phage-assisted continuous evolution are provided herein. These include strategies, systems, methods, reagents, and kits allowing for stringency modulation to evolve weakly active or inactive biomolecule variants, negative selection of undesired properties, and / or positive selection of desired properties.

The invention discloses a bispecific nano antibody targeting CXCL1 and PD-L1 and application thereof, and belongs to the technical field of antibody engineering and bioengineering. The bispecific nano-antibody targeting CXCL1 and PD-L1 comprises at least one nano-antibody for recognizing a target CXCL1 and a nano-antibody for recognizing a target PD-L1, and nano-antibody monomers are connected through a linker. The bispecific nano-antibody targeting PD-L1 / CXCL1 provided by the invention has a unique structure, so that the bispecific nano-antibody targeting PD-L1 / CXCL1 has specific recognition and binding capacities to CXCL1 and PD-L1, the affinity to CXCL1 reaches 1.08 nM, the affinity to PD-L1 reaches 1.09 nM, drug resistance of a patient to antibody treatment caused by immune escape can be effectively prevented, and the bispecific nano-antibody targeting PD-L1 / CXCL1 is suitable for more patients and has good clinical application prospects. And a new thought is provided for CRC treatment.

The invention discloses an anti-TP53 mutation T cell receptor and application thereof, the T cell receptor comprises specific alpha chain and beta chain variable domains, and the complementary determining region (CDR) sequence is shown as SEQ ID NO: 9-14. The TCR has the core advantage that the TCR has excellent broad-spectrum recognition capability, can target six different amino acid substitutions (A, G, I, N, S and T) at the R249 site, and effectively deals with tumor heterogeneity and mutation difference between patients. Aiming at high-frequency HLA-B * 07: 02 alleles in people, the TCR lays a foundation for developing TCR-T cell therapy covering a wide range of people, and has great clinical application value and market potential in treatment of various solid tumors carrying TP53 R249 hotspot mutation, such as liver cancer.

The present disclosure provides an antibody or antigen-binding fragment thereof that specifically binds to human cMET. The disclosure also provides polynucleotides encoding the antibodies or antigen binding fragments thereof, vectors comprising the polynucleotides, host cells comprising the vectors, methods of generating the antibodies, and compositions comprising the antibodies.

The invention provides a monoclonal antibody of a targeted human folate receptor alpha (FR1). Specifically, a mouse is immunized through FR1 recombinant protein, and a monoclonal antibody with high affinity to FR1 is screened out. In addition, the invention also provides an amino acid sequence of the monoclonal antibody, nucleic acid containing the sequence, a carrier or a conjugate containing the nucleic acid, and application of the monoclonal antibody in FR1 overexpressed tumors / cancers.

The disclosure provides, for example, double stranded DNA (dsDNA) molecules comprising one or more chemically modified nucleobases. In some embodiments, the dsDNA molecule is circular and comprises a first strand and a second strand, wherein the first strand comprises one or more chemically modified nucleobases, and the second strand is free of chemically modified nucleobases. In some embodiments, the dsDNA molecule comprises a promoter sequence and an effector sequence that encodes an effector.

The invention discloses a polyphosphate kinase mutant, a biological material, a catalyst and application, and belongs to the field of protein engineering, and compared with a wild enzyme, the mutant has one or more amino acid residue mutations in the 307th site, the 311th site, the 320th site, the 408th site, the 419th site and the 431th site. The mutant disclosed by the invention is obtained by combining a semi-rational design strategy with active site region prediction and mutation library design screening, and the enzyme activity and the relative yield are remarkably improved: the efficiency of catalytic synthesis of GTP by using the preferable mutant Y431P / K311F / W408A with GMP as a substrate is 2.6 times that of a wild type enzyme, and the relative yield is improved by 150%; the efficiency of the optimized mutant Y431P / K311F / R419L for catalytically synthesizing GTP by taking GDP as a substrate is 1.9 times that of a wild enzyme, and the relative yield is increased by 82%. The key mutant obtained by screening can realize efficient synthesis from cheap NDP or NMP to high-value NTP, and has an excellent industrial application prospect.

The invention provides an H1N1 broad-spectrum mRNA (messenger ribonucleic acid) vaccine taking HA (hemagglutinin) and NA (neuraminidase) protein tandem as a target and a preparation method of the H1N1 broad-spectrum mRNA vaccine, and relates to the technical field of vaccine preparation, the amino acid sequence of the mRNA vaccine is shown as SEQ ID NO.1, and the nucleotide sequence of the mRNA vaccine is shown as SEQ ID NO.2, namely, the mRNA vaccine is obtained by connecting GGGSGGSGGGSGGGGS with conserved amino acid sequences of hemagglutinin HA and neuraminidase NA. The defects in the prior art are overcome, and the broad spectrum and the protection effect of the vaccine are improved, so that the vaccine can better cope with the immune escape of the latest variant in the future.

The invention relates to the technical field of biological medicine, in particular to an antibody aiming at staphylococcus aureus enterotoxin B and application thereof. A heavy chain variable region of the antibody provided by the invention comprises CDR sequences as shown in SEQ ID NO.1-3, and a light chain variable region of the antibody comprises CDR sequences as shown in SEQ ID NO.9-11. The antibody has high affinity, can specifically bind to staphylococcus aureus SEB, can block binding of SEB and MHC II / TCR, significantly inhibits cytokine storm, shows a dose-dependent protection effect in an MRSA systemic infection model, and can be used for preparing an MRSA systemic infection model. The monoclonal antibody can be used for treating, preventing or diagnosing the infection of the staphylococcus aureus, provides a solution of non-antibiotic therapy for SEB poisoning and drug-resistant staphylococcus aureus infection, and has important clinical and public health values.

The invention discloses a novel microecological preparation composition for preventing and treating feline plague and application of the novel microecological preparation composition, and belongs to the technical field of microecological preparations. The recombinant lactobacillus plantarum NC8-VP2 constructed by a genetic engineering means can efficiently express the feline plague VP2 protein, and finally, a protective antigen expressed by the recombinant lactobacillus plantarum efficiently stimulates an organism to generate a specific antibody, so that the feline plague is prevented, and the use of chemical drugs is reduced. Recombinant escherichia coli BL21-pET32a-MLT constructed by the research can secrete and express melittin (MLT), large-scale production of melittin is realized, melittin and recombinant lactobacillus plantarum NC8-VP2 are further taken as active ingredients, a novel microecological preparation composition for preventing and controlling feline plague is developed and prepared, the morbidity of feline plague can be reduced, clinical symptoms of sick animals can be improved, and the novel microecological preparation composition has a good application prospect. Therefore, the compound has a relatively good application prospect in the aspect of preventing and treating feline plague.

Provided are a fusion protein (for example, in the form of a bispecific antibody) binding to CD235a and CD3, a preparation method therefor, and a related use thereof.