33 results about "Chain gene" patented technology

The invention provides an establishment method of a transgenic mouse, and the method comprises the following steps: a transgenic mouse MHC chimeric gene is introduced into an endogenous B2m gene locus of the mouse, the MHC chimeric gene comprises a human B2M gene and a chimeric alpha-chain gene, and the chimeric alpha-chain gene comprises a human alpha1 region, a human alpha2 region and a mouse alpha3 region; the chimeric alpha-chain gene further comprises a mouse transmembrane region and a mouse intracellular region. The transgenic mouse disclosed by the invention can be applied to tumor specific antibody detection, in-vivo evaluation of vaccines, and in-vivo screening of TCR-T and other drugs.

The invention discloses application of a ZmFAH1 gene in regulating and controlling low temperature resistance of corn in a germination period, a related molecular marker and application of the molecular marker. The invention firstly provides the high-low-temperature-resistant ZmFAH1 gene, and the relative expression quantity of the high-low-temperature-resistant ZmFAH1 gene is obviously higher than that of a low-temperature sensitive material in embryos in a germination stage, buds in a bud stage, roots in the bud stage and roots in a seedling stage; the ZmFAH1 gene can significantly improve the low temperature resistance of corn in the germination period, and also can enhance the low temperature resistance of corn in the germination period and the seedling period. The invention further provides a molecular marker DNCAPS708 which is located at the 708 -710 sites of the CDS region of the negative chain ZmFAH1 gene of the first chromosome of the corn, and three deleted basic groups are GAA / <->. The research finds that the overexpression of the ZmFAH1 gene can effectively improve the low temperature resistance of the corn, which is of great significance to the molecular biology mechanism research of the low temperature resistance function of the corn.

The present invention discloses a recombinant collagen with enhanced skin regeneration ability, a preparation method thereof and an application thereof. The present invention provides a recombinant type III collagen based on the gene sequence of human type III collagen α1 chain, and the encoding nucleic acid molecule sequence thereof is shown in SEQ NO.1. The recombinant type III collagen is expressed by high-density fermentation of Pichia pastoris GS115 strain, combined with optimized cation exchange chromatography and anion exchange chromatography purification processes, to achieve efficient preparation with a protein purity of 99.2% and a yield of 13.8 g / L; the recombinant type III collagen is highly consistent with natural collagen, and can significantly promote skin regeneration, collagen fiber deposition and wound repair.

The present invention discloses a genetically modified rodent whose immunoglobulin light chain locus is modified so as to comprise a single rearranged human immunoglobulin light chain gene V / J gene upstream of a mouse Kappa light chain locus constant region, a mouse variable region sequence cannot bind to an endogenous constant region to form a complete mouse light chain antibody, the rodents are capable of normally reproducing and producing human light chain-containing antibodies. The invention also provides a method of making the genetically modified rodent and a use of the rodent.

Phospholamban (PLN) is a critical regulator of calcium cyclin and contractility in the heart. The deletion of Arginine 14 of the phospholamban gene (R14del) is associated with the pathogenesis of an inherited form of cardiomyopathy with prominent arrhythmias. Although the genetic etiology is well defined, there are currently no therapies for this rare disease. This disclosure provides an allele-specific silencing approach by interfering RNA (RNAi) to reduce the expression levels of the R14del allele of the PLN gene.

To provide a humanized rodent that expresses a heavy chain containing a VL domain. [Solution] A non-human animal, tissue, cell, and genetic material is provided which includes a modification of an endogenous non-human heavy chain immunoglobulin sequence and contains functional ADAM6 activity in a rodent (e.g., mouse), wherein the non-human animal rearranges the human immunoglobulin light chain gene segment in relation to the heavy chain constant domain and expresses an immunoglobulin-like molecule which contains a human immunoglobulin light chain variable domain fused to the heavy chain constant domain and is a cognitive of a human immunoglobulin light chain variable domain fused to the light chain constant domain.

This invention provides an organ fibrosis-related micropeptide, OFMP, and its applications. This type of micropeptide, OFMP, is a translational product with endogenous biological activity and stable existence, encoded by an open reading frame (ORF) in a long non-coding RNA. It is the first time that the long non-coding RNA encoding the micropeptide has been found to be significantly related to organ fibrosis and can encode the micropeptide. Synthetic peptides prepared based on the aforementioned OFMP micropeptide show significant inhibitory effects on several key indicators of organ fibrosis, including type I collagen α1 chain gene, α-smooth muscle actin, transforming growth factor β1, and connective tissue growth factor. This indicates that the OFMP micropeptide, the nucleotide sequence (ORF) encoding the micropeptide, and the long non-coding RNA sequence containing the nucleotide sequence have important clinical diagnostic and therapeutic value for organ fibrosis.

Mice are provided that comprise a reduction or deletion of ADAM6 activity from an endogenous ADAM6 locus, or that lack an endogenous locus encoding a mouse ADAM6 protein, wherein the mice comprise a sequence encoding an ADAM6 or ortholog or homolog or fragment thereof that is functional in a male mouse. In one embodiment, the sequence is an ectopic ADAM6 sequence or a sequence that confers upon a male mouse the ability to generate offspring by mating. Mice and cells with genetically modified immunoglobulin heavy chain loci that comprise an ectopic nucleotide sequence encoding a mouse ADAM6 or functional fragment or homolog or ortholog thereof are also provided.

Provided herein is a transgenic chicken comprising a genome having an endogenous immunoglobulin heavy chain gene in which the IgY CH1 coding sequence is functionally deleted. In certain embodiments, in B cells of the chicken the endogenous immunoglobulin heavy chain gene comprises: (a) a functional immunoglobulin heavy chain gene comprising, in operable linkage, a nucleic acid encoding an antibody heavy chain variable domain and a constant region coding sequence in which the IgY CH1 coding sequence is functionally deleted; and (b) a plurality of pseudogenes that are operably linked to said functional immunoglobulin heavy chain gene and that donate, by gene conversion, nucleotide sequence to the nucleic acid encoding the variable domain of (a), wherein the pseudogenes are upstream of the nucleic acid encoding an antibody heavy chain variable domain of (a).

This disclosure provides, among other things, a transgenic animal that uses gene conversion for antibody diversification, comprising B cells in which the endogenous immunoglobulin heavy chain locus comprises: (a) a functional immunoglobulin heavy chain gene comprising a nucleic acid encoding an autonomous heavy chain (AHC) variable domain; and (b) a plurality of pseudogenes that are operably linked to said functional immunoglobulin heavy chain gene and that donate, by gene conversion, nucleotide sequence to the nucleic acid encoding the AHC variable domain of (a), wherein the pseudogenes are upstream or downstream of the functional immunoglobulin heavy chain gene.

This disclosure provides, among other things, a transgenic chicken. In some embodiments, the transgenic chicken comprises B cells in which the endogenous immunoglobulin heavy chain locus comprises: (a) a functional immunoglobulin heavy chain gene comprising a nucleic acid encoding a heavy chain variable domain in which the CDR3 is in the range of 30-60 amino acids in length and comprises at least 2 cysteine residues; and (b) a plurality of pseudogenes that are operably linked to said functional immunoglobulin heavy chain gene and that donate, by gene conversion, nucleotide sequence to the nucleic acid encoding the heavy chain variable domain of (a), wherein the pseudogenes are upstream or downstream of the functional immunoglobulin heavy chain gene.

This disclosure provides, among other things, strategies for minimizing antibody diversification in a transgenic animal that uses gene conversion for antibody diversification. In some embodiments, the animal may comprise a genome comprising an endogenous immunoglobulin light chain locus comprising: (a) a functional immunoglobulin light chain gene comprising a nucleic acid encoding a light chain variable region; and (b) a plurality of pseudogenes that are operably linked to the functional immunoglobulin light chain gene and that donate, by gene conversion, nucleotide sequence to the nucleic acid encoding a light chain variable region, wherein the pseudogenes are upstream or downstream of the functional immunoglobulin light chain gene and encode the same amino acid sequence as the light chain variable region of the functional immunoglobulin light chain gene of (a). In other embodiments, the locus may have a tandem array of coding sequences for the light chain.

This disclosure provides, among other things, a transgenic chicken. In some embodiments, the transgenic chicken comprises B cells in which the endogenous immunoglobulin heavy chain locus comprises: (a) a functional immunoglobulin heavy chain gene comprising a nucleic acid encoding a heavy chain variable domain in which the CDR3 is in the range of 30-60 amino acids in length and comprises at least 2 cysteine residues; and (b) a plurality of pseudogenes that are operably linked to said functional immunoglobulin heavy chain gene and that donate, by gene conversion, nucleotide sequence to the nucleic acid encoding the heavy chain variable domain of (a), wherein the pseudogenes are upstream or downstream of the functional immunoglobulin heavy chain gene.

The present invention provides, among other things, a transgenic chicken. In some embodiments, the transgenic chicken comprises a B cell having an endogenous immunoglobulin heavy chain locus comprising: (a) a functional immunoglobulin heavy chain gene comprising a nucleic acid encoding a heavy chain variable domain having a length of CDR3 in the range of 30 to 60 amino acids and containing at least two cysteine ​​residues; and (b) a plurality of pseudogenes operably linked to the functional immunoglobulin heavy chain gene and, by gene conversion, providing a nucleotide sequence to the nucleic acid encoding the heavy chain variable domain of (a), wherein the pseudogenes are located upstream or downstream of the functional immunoglobulin heavy chain gene.

The present invention provides: a composition which is for increasing the efficiency of genome editing with a single-strand-form editing polynucleotide that is capable of modifying a target site in a double-stranded genome DNA, the composition containing at least one protein selected from the group consisting of Hfq, DnaK, and variants thereof, or a nucleic acid comprising a base sequence that encodes the protein; a genome editing kit comprising the composition and an editing polynucleotide; a method for modifying a target site in a double-stranded genome DNA of a cell or a living organism, the method comprising a step of using the genome editing kit to treat a cell or living organism; and a method comprising said step, for producing a cell or living organism in which a target site in a double-stranded genome DNA has been modified.

To provide a hybrid light chain mouse.SOLUTION: There is provided a mouse genetically modified to appear a Homo sapiens λ variable (hVλ) sequence (containing a mouse for appearing the hVλ sequence from an endogenous mouse λ light chain gene locus, a mouse for appearing an hVλ sequence from an endogenous mouse κ light chain gene locus, and a mouse for appearing hVλ sequence from a transducing gene or an episome). Here, the hVλ sequence is connected to a mouse steady sequence. There is provided a mouse for becoming an origin of the Homo sapiens λ variable sequence which has been varied into a body cell variation useful for preparing an antigen binding protein. There are provided a composition and a method for preparing an antigen binding protein containing a human λ variable sequence containing a human antibody.SELECTED DRAWING: Figure 7B

A composition for increasing the efficiency of genome editing with a single-strand-form editing polynucleotide capable of modifying a target site in a double-stranded genome DNA. The composition includes at least one protein selected from Hfq, DnaK, and variants thereof, or a nucleic acid having a base sequence that encodes the protein. A genome editing kit including the composition and an editing polynucleotide. A method for modifying a target site in a double-stranded genome DNA of a cell or a living organism including treating a cell or living organism with the genome editing kit. A method for producing a cell or living organism in which a target site in a double-stranded genome DNA has been modified.

The invention discloses a cell line capable of stably expressing a porcine PIGR SC fragment and a J chain as well as a construction method and application of the cell line, and belongs to the technical field of gene engineering. The construction method comprises the following steps: inserting coding sequences of an SC fragment gene and a J-chain gene of the porcine PIgR into lentiviral vector plasmids to obtain an SC fragment and J-chain overexpressed lentiviral vector plasmid of the PIgR; carrying out co-transfection on the lentivirus vector plasmid, PPSAX2 and PMD2. G plasmids, so as to obtain a recombinant lentivirus; and infecting cells with the recombinant lentivirus to obtain positive cells, and carrying out enlarged culture to obtain the SC fragment and J chain cell line for stably expressing the swine PIGR. According to the invention, a cell line for stably expressing the SC fragment and the J chain of the porcine PIgR is constructed, so that the SIgA antibody can be rapidly and efficiently prepared by a gene engineering technology.

This disclosure provides, among other things, strategies for minimizing antibody diversification in a transgenic animal that uses gene conversion for antibody diversification. In some embodiments, the animal may comprise a genome comprising an endogenous immunoglobulin light chain locus comprising: (a) a functional immunoglobulin light chain gene comprising a nucleic acid encoding a light chain variable region; and (b) a plurality of pseudogenes that are operably linked to the functional immunoglobulin light chain gene and that donate, by gene conversion, nucleotide sequence to the nucleic acid encoding a light chain variable region, wherein the pseudogenes are upstream or downstream of the functional immunoglobulin light chain gene and encode the same amino acid sequence as the light chain variable region of the functional immunoglobulin light chain gene of (a). In other embodiments, the locus may have a tandem array of coding sequences for the light chain.

Genetically modified non-human animals are provided that express an immunoglobulin variable domain that comprises at least one histidine, wherein the at least one histidine is encoded by a substitution of a non-histidine codon in the germline of the animal with a histidine codon, or the insertion of a histidine codon in a germline immunoglobulin nucleic acid sequence. Immunoglobulin genes comprising histidines in one or more CDRs, in an N-terminal region, and / or in a loop 4 region are also provided. Immunoglobulin variable domains comprising one or more histidines (e.g., histidine clusters) substituted for non-antigen-binding non-histidine residues. Non-human animals that are progeny of animals comprising modified heavy chain variable loci (V, D, J segments), modified light chain variable loci (V, J segments), and rearranged germline light chain genes (VJ sequences) are also provided. Non-human animals that make immunoglobulin domains that bind antigens in a pH-sensitive manner are provided.

The invention relates to the technical field of detection experiments, in particular to an automatic scheduling method and system for assembly line experiments and related equipment, and the method comprises the steps: randomly generating a full-permutation array as an initial population, the full-permutation array representing different detection sequences of each detection experiment; taking the initial population as a parent, performing crossover variation by introducing linkage gene probability in biology, determining a target population, and obtaining total detection duration of each individual in the target population; and taking the individual corresponding to the minimum value in the total detection duration as a target individual, and detecting each detection experiment according to the detection sequence corresponding to the target individual. According to the method, the tasks are scheduled and arranged by adopting the genetic algorithm, and the detection efficiency is optimized.

The present application discloses silk improved by a spider silk protein gene and a method for using a spider silk protein gene to improve silk. The silk improved by the spider silk protein gene comprises a silk protein light chain. The silk protein light chain is linked to a spider silk protein which is formed by fusion expression with the silk fibroin light chain. An amino acid sequence of the spider silk protein comprises a repeat region sequence having at least one spider silk protein repeat unit sequence. The present application achieves the expression of Nephila pilipes pyriform silk in Bombyx mori silk fibroin, implements formation of an intact spider silk protein on the silk light chain, and expands sites for Bombyx mori silk gland gene editing. Light-chain gene edited Bombyx mori and a heavy-chain gene edited silkworm can be hybridized, thereby providing technical support for further selective breeding and optimization of fiber performance.

The present invention relates to seven novel neutralizing human monoclonal antibodies (mAbs) THSC20.HVTR04, THSC20.HVTR06, THSC20.HVTR11, THSC20.HVTR26 THSC20.HVTR39, THSC20.HVTR55 and THSC20.HVTR88 and their nucleotide sequences isolated from a convalescent individual of Indian origin by antigen (RBD)-specific single B cell sorting and cloning of variable heavy and light IgG chain genes. The isolated mAbs demonstrate neutralization of wild type Wuhan strain and the following variants of concern: South African variant of concern (B.1.351), UK variant of concern (B.1.1.7), Brazilian variant of concern (PI), Delta (B.1.617.2) and Omicron (B.1.1.529) with exception of THSC20.HVTR39 unable to neutralize Gamma (P1). Of these THSC20.HVTR04 is able to potently neutralize Omicron BA.2 and BA.4 / BA.5, THSC20.HVTR06 is able to neutralize Omicron BA.1, BA.2 and BA.5 with low potency, THSC20.HVTR11 potently neutralizes Omicron BA.1 and BA.2 and THSC20.HVTR26 neutralizes Omicron BA. 1 only with moderate potency. The present invention also discloses the binding affinity of the neutralizing mAbs to the receptor binding domain (RBD) representing Wuhan isolate (wild type). The present invention also, discloses the use of neutralizing monoclonal antibodies (mAbs) against SARS-CoV-2 for its diagnostic, prognostic, preventive and therapeutic purposes.

Provided are an organ fibrosis-related micropeptide (OFMP) and a use thereof. The OFMP is a stable translation product coded by an open reading frame (ORF) in a long non-coding RNA and having endogenous biological activity. It is found for the first time that the long non-coding RNA coding the micropeptide has an important relationship with organ fibrosis and can code the micropeptide. A synthetic peptide prepared from the OFMP has a significant inhibitory effect on a plurality of organ fibrosis key indicators, comprising a collagen type I α1 chain gene, α-smooth muscle actin, a transforming growth factor β1, and a connective tissue growth factor, indicating that the OFMP, a nucleotide sequence (ORF) for coding the micropeptide, and a long non-coding RNA sequence containing the nucleotide sequence have important clinical diagnosis and clinical treatment value on organ fibrosis.

The invention provides a preparation method of an ictalurus punctatus IgM antibody based on a prokaryotic expression system, and relates to the field of aquatic animal immunodetection and antibody preparation, and the preparation method comprises the following steps: designing a specific primer based on an ictalurus punctatus IgM protein gene sequence in GenBank, and carrying out antigen determinant truncation; the method comprises the following steps: extracting liver RNA (Ribonucleic Acid) of channel catfish by a Trizol method, reversely transcribing the liver RNA into cDNA (Complementary Deoxyribonucleic Acid), and cloning an IgM The method comprises the following steps: constructing a pET-30a-IgM recombinant expression vector, transforming Escherichia coli Rosetta competent cells, and inducing protein expression; purifying the truncated IgM protein by adopting the steps of ultrasonication, affinity chromatography and dialysis concentration; a New Zealand white rabbit is immunized to prepare antiserum, and the titer is detected through ELISA. According to the method, efficient and low-cost preparation of the IgM protein is realized by utilizing a prokaryotic expression system, the period is only several weeks, high-purity protein of milligram-grade to gram-grade can be obtained, the batch stability is good, related biological safety risks of animal serum are avoided, and a key basis is provided for development of an ictalurus punctatus ELISA (enzyme-linked immunosorbent assay) kit, immunohistochemistry and B lymphocyte research.

Adeno-associated virus (AAV) expression vectors for the treatment of Charcot-Marie-Tooth disease are provided. The AAV expression vectors can have at least one neurofilament light chain gene (NEFL)-targeting shRNA operably linked to a first promoter and a NEFL cDNA operably linked to a second promoter. Pharmaceutical compositions comprising AAV expression vectors are also provided, as are methods of treating Charcot-Marie-Tooth disease type 2E.

The present invention provides a method for producing a circular vector that includes a long-chain gene cluster without mutations, the method involving ligating, in one pod and in one step in the presence of an exonuclease, a DNA polymerase, a DNA ligase, and a master mix, a linear vector and three or more DNA fragments having an overlap of 40-60 bp on both ends and constituting a long-chain gene cluster.

According to the dsRNA of the migratory locust myosin heavy chain gene and the application of the dsRNA, the dsRNA for synthesizing interference sequence fragments of the migratory locust myosin heavy chain gene is injected into migratory locust bodies, so that the mRNA expression of the migratory locust myosin heavy chain gene can be specifically silenced, the excavation frequency is obviously reduced when insects lay eggs, and the excavation depth is obviously reduced; a large number of eggs are retained in the body, so that insects cannot normally lay eggs, and the purpose of biological control is achieved.

To provide a heterologous recombinant baculovirus (rBV) expression system for production of foreign heterologous proteins in insect cells.SOLUTION: This system comprises: a recombinant baculovirus backbone genome with a deletion in the cathepsin gene into which foreign gene cassettes can be integrated; and an insect cell which can be infected with the Δv-cath-rBV, and in which foreign proteins and / or viral vectors or particles are expressed.SELECTED DRAWING: Figure 1A

Disclosed is a method for assaying a sample of viral particles that contain single-stranded DNA genome for strand specificity.In an embodiment, the method includes: synthesizing a synthetic DNA strand that is complementary to the single-stranded DNA genome by incorporating modified bases to obtain labeled double-stranded DNA products; purifying the labeled double-stranded DNA products; sequencing the purified double-stranded DNA products on a sequencer; and determining the specificity of the viral single-stranded DNA genome based on detecting the modified bases of the synthetic DNA strand.