12 results about "Plasmid Vector" patented technology

A method for improving the replication of covalent closed circular plasmids is provided. [Solution] The method comprises the step of preparing a covalent closed circular plasmid having a Pol I-dependent origin of replication and an insert containing a structured DNA sequence selected from the group consisting of a reverse repeat sequence, a directional repeat sequence, a homopolymer repeat sequence, a eukaryotic origin of replication, or a eukaryotic promoter enhancer sequence, wherein the structured DNA sequence is located at a distance of less than 1000 bp from the Pol I-dependent origin of replication in the direction of replication. The method also comprises the step of modifying the covalent closed circular recombinant molecule so that the Pol I-dependent origin of replication is replaced with a Pol III-dependent origin of replication, thereby improving the replication of the covalent closed circular plasmid with the resulting Pol III-dependent origin of replication. A covalent closed circular recombinant DNA molecule without antibiotic markers is also provided.

This invention provides a method for constructing a visualized mouse model characterizing hepatocyte necrosis and apoptosis, belonging to the field of biomedical technology. The invention involves disassembling firefly luciferase (Fluc) into inactive N-terminal (Nluc) and C-terminal (Cluc) fragments, which are then fused and expressed at specific locations on the MLKL molecule, forming two independent plasmid vectors, MLKL-Nluc and MLKL-Cluc. These two plasmid vectors are then injected into mouse hepatocytes via tail vein high-pressure hydrodynamic injection to construct the visualized mouse model characterizing hepatocyte necrosis and apoptosis. This invention allows for non-invasive and dynamic monitoring of the execution process of necrosis and apoptosis—MLKL oligomerization—in animal models, providing a powerful and specific tool that fills the technological gap in real-time visual monitoring of necrosis and apoptosis in pan-apoptotic research.

The application discloses a pSY1A plasmid vector for a wheat germ cell-free expression system, which is constructed by introducing an ampicillin resistance gene, an Ori replicon, a T7 promoter, a translation enhancer, a MCS (multiple cloning site), a 3'-UTR sequence and a T7 terminator sequence on the basis of part of a sequence of an E. coli plasmid pUC19. The pSY1A plasmid vector has a MCS into which an exogenous gene can be inserted, and the MCS and a transcription assembly constitute a transcription module, so that the transcription module can be preserved by preserving the plasmid, and in-vitro transcription of the wheat germ cell-free expression system becomes more convenient and faster.

The application discloses an antiepileptic preparation preparation method and application of a micro ribonucleic acid 145b targeting agent, and relates to the technical field of biological medicine. The method comprises the following steps: constructing a neuron-specific recombinant adeno-associated virus plasmid vector, wherein the vector comprises a neuron-specific promoter hSyn, a reporter gene, an enhancer, and a functional nucleic acid sequence for targeting and inhibiting micro ribonucleic acid 145b; using an adeno-associated virus packaging system to transfect the vector to a packaging cell for virus packaging; collecting a lysis solution and separating out a recombinant adeno-associated virus particle through a purification process; and resuspending the virus particle to obtain a preparation. The application specifically down-regulates the miR-145b level in hippocampal neurons, releases the inhibition of the Kcnq3 potassium ion channel, further up-regulates the Kcnq3 expression, restores the M current, and reduces the neuron excitability from a molecular mechanism.

This invention discloses a method for establishing GLI1 deacetylated or pseudoacylated mutants, relating to the field of molecular biology technology; comprising: S1: identifying GLI1 protein acetylation sites; S2: amplifying GLI1 deacetylated or pseudoacylated mutant gene fragments; S3: amplifying plasmid vectors; S4: ligating the GLI1 deacetylated or pseudoacylated gene fragments to the plasmid vectors; S5: verifying GLI1 deacetylated or pseudoacylated mutant protein expression using immunofluorescence and Western blot (WB). The method for establishing GLI1 deacetylated or pseudoacylated mutants of this invention can stably maintain the GLI1 deacetylated or pseudoacylated state.

“Plasmid vector for DNA delivery, vaccine and / or therapeutic compositions and uses” The present invention relates to a plasmid vector for DNA delivery using lactic acid bacteria, which can be used for both DNA vaccines and gene therapy. The vector proposed in the present invention has a theta origin of replication, which confers greater stability; in addition to an expression cassette containing a multiple cloning site and a eukaryotic promoter, which allows the vector to have a wide variety of host cells. The vaccine and / or therapeutic compositions comprising the vector can be used for the treatment and / or prophylaxis of various diseases.

The invention discloses a red light reversible regulation gene expression system and application, the system comprises three plasmid vectors, respectively a plasmid pPA formed by fusing a PUF protein and a phytochrome PhyA, a plasmid pFE formed by fusing a shuttle protein FHY1 and eIF4E, and a downstream targeting regulation plasmid pNG formed by an NRE target and a downstream gene of interest GOI; according to the invention, the binding characteristic of RNA binding protein PUF and target NRE thereof is utilized, and the characteristic of dimerization of PhyA protein and ligand FHY1 thereof under red light is fused, so that a gene expression activation system capable of bidirectional light regulation is constructed, the PUF protein continuously and stably inhibits expression of downstream genes of interest, background leakage of the system is reduced from the source, and the expression of the downstream genes of interest is inhibited. Expression of downstream interested genes can be caused only under irradiation of red light, the expression can be rapidly closed by far-red light, and the expression of the genes can be accurately controlled through light; the system can be applied to tumor treatment.

The present invention relates to an isolated polypeptide comprising an amino acid sequence having aminoacylase activity and having at least 87% sequence identity with the amino acid sequence shown in SEQ ID NO: 1 over its entire length. The present invention further relates to an isolated nucleic acid molecule comprising a nucleotide sequence encoding aminoacylase, a plasmid vector comprising the nucleic acid molecule, a recombinant host cell comprising the isolated nucleic acid molecule or vector, and a method for producing the aminoacylase. The present invention also encompasses the use of the aminoacylase according to the present invention for N-acylation of amino acids or salts thereof, and a method for producing N-acylamino acids or salts thereof using the aminoacylase. Furthermore, the present invention relates to the obtained N-acylamino acids, compositions comprising the same, and the use of the produced N-acylamino acids in cosmetic products, home care products, or professional products.

The application provides a cysteine-strengthened recombinant spider silk and a preparation method and application thereof. An amino acid sequence of the recombinant spider silk protein has the sequence shown in SEQ ID NO: 6. A polynucleotide encoding the amino acid sequence of the protein has the following structure: a 5' end of a cysteine-modified MaSp2 gene of the major ampullate gland of the spider Argiope bruennichi is connected with an N-domain, and a 3' end is connected with a C-domain; the gene is inserted into a pET-28a plasmid vector to obtain a recombinant plasmid, and the recombinant plasmid is introduced into E. coli to express the recombinant spider silk protein. The recombinant spider silk prepared by wet spinning using the recombinant spider silk protein has a dense beta-sheet crystal structure, excellent mechanical properties and wet strength, and opens up a prospect for industrial application of the recombinant spider silk.

OCR06WT The current invention involves guide RNA (gRNA) for use in modification. Genetic precursors of host microorganisms and methods for genetic modification of host microorganism species. Escherichiacoli was used to induce the expression of target genes using the CRISPR / CASS system. Such genetic modification involves introducing a target gene into a specific intergene region. of the host microorganisms using a CRISPR / Case system in which the CRISPR / Case system is comprised of (5) A plasmid vector guide RNA (gRNA plasmid) containing specific guide RNA (gRNA). To modify the desired position on the specific intergene region (i) plasmid vector redcas-CM. (REDCas-Cmplasmid) and Gii) prototypes for CAMP repair consisting of nucleotide sequences. Target gene removal and degradation of plasmid welter-guided RNA and plasmid vector redcast. CM is released from genetically modified microorganisms containing a guide RNA nucleotide sequence. Choose one or more nucleotide sequences from 1-38 (SEQIDNO:1-38). Combined

This invention belongs to the fields of biomedicine and in vitro diagnostics, specifically relating to a specific antibody against porcine epidemic diarrhea virus (PEDV), a nucleic acid molecule, a plasmid vector combination, and a eukaryotic cell expression vector. The amino acid sequence of the specific antibody of this invention consists of two dominant antigenic epitopes of the PEDV N protein tandemly. Using E. coli-preferred codons, the amino acid sequence of this recombinant protein is converted into the corresponding nucleotide sequence, thereby increasing the expression level of the recombinant protein in E. coli. This invention also involves immunizing mice with this recombinant protein, sorting B lymphocytes that specifically bind to the recombinant protein using flow cytometry, amplifying the heavy and light chain variable regions of the B lymphocyte antibody using single-cell PCR, constructing a complete mouse IgG antibody sequence recombinant expression vector from the obtained sequence, expressing the monoclonal antibody by transiently transfecting HEK293F cells, purifying the monoclonal antibody and labeling it with colloidal gold particles, and determining the optimal monoclonal antibody pairing combination through orthogonal experiments.