17 results about "Plasmidome" patented technology

This invention relates to a method and its application for determining the plasmid content in a quadrivalent avian influenza DNA vaccine. Belonging to the field of molecular biology, this invention aims to provide a TaqMan quantitative real-time PCR method capable of quantitatively determining the content of each plasmid component in a quadrivalent DNA vaccine. Specifically, this invention provides a primer and probe composition for detecting the content of individual plasmids in a quadrivalent avian influenza (H5+H7) DNA vaccine based on TaqMan quantitative real-time PCR. The composition includes primers with nucleotide sequences shown in SEQ ID NO: 5-6, 8-9, 11-12, and 14-15, and TaqMan probes with nucleotide sequences shown in SEQ ID NO: 7, 10, 13, and 16. This primer and probe composition can quantitatively determine the content of each plasmid component in the quadrivalent DNA vaccine, exhibiting good specificity and reproducibility.

The invention relates to a bombyx mori DNA 6mA methylation modification system and application thereof, and belongs to the technical field of biology. According to the invention, a bombyx mori specific DNA 6mA methylation modification system is constructed, and the system is composed of a pdCas9-BmMETTL4-EGFP plasmid and a pU6-gRNA plasmid; an inactivated dCas9 gene is optimally designed based on a silkworm codon, and a 6mA modification function is realized by fusing BmMETTL4 methyltransferase and a fluorescent label; and guiding the system to target a target gene region through the specific gRNA. The system can significantly enhance the 6mA methylation level in a target gene area, effectively inhibit the expression of BmPSB1 and BmMETTL4 genes, and show functions in the regulation and control of cell mitosis. The method breaks through the technical bottleneck of insect epigenetic editing, provides a core technical tool for precise regulation and function research of bombyx mori genes, and has the characteristics of high efficiency, targeting property and application universality.

Provided in the present disclosure is a method for using a four-plasmid system to prepare modified immune effector cells. The method comprises: forming a lentivirus by using four plasmids within 293T cells, extracting and obtaining the lentivirus, then transfecting immune effector cells by using the lentivirus, and expressing a chimeric antigen receptor. Also provided in the present disclosure is a use of the immune effector cell obtained by using the described method and of a composition containing the immune effector cell.

The present invention discloses a recombinant Yarrowia lipolytica and its application in the production of erythritol, belonging to the field of bioengineering technology. The Ku70 gene has been deleted in the recombinant Yarrowia lipolytica genome of the present invention, and the GUT1, GUT2, TKL1 and TAL1 genes are overexpressed using the plasmid pINA1269 combination. The present invention provides a recombinant Yarrowia lipolytica Y-11 / pINA1269-GUT1-TKL1-GUT2-TAL1 that can produce high erythritol. The recombinant Yarrowia lipolytica is inoculated into a 5L fermenter and fermented for 120 hours using crude glycerol as a carbon source. The concentration of erythritol in the fermentation broth can be as high as 110±1.1g / L, and the conversion rate can be as high as 61%.

The invention relates to a method for determining the plasmid content in an avian influenza tetravalent DNA vaccine and application. The invention belongs to the field of molecular biology, and aims to provide a TaqMan fluorescent quantitative PCR (Polymerase Chain Reaction) method capable of quantitatively determining the content of each plasmid component in a tetravalent DNA vaccine. Specifically, the invention provides a primer and probe composition for detecting the content of a single plasmid in the avian influenza (H5 + H7) tetravalent DNA vaccine based on TaqMan fluorescent quantitative PCR, and the primer and probe composition comprises primers with nucleotide sequences as shown in SEQ ID NO: 5-6, 8-9, 11-12 and 14-15, and TaqMan probes with nucleotide sequences as shown in SEQ ID NO: 7, 10, 13 and 16. The primer and probe composition can quantitatively determine the content of each plasmid component in the tetravalent DNA vaccine, and has good specificity and repeatability.

The present application relates to the field of genetic engineering, in particular to a sgRNA group targeting DYA gene, a primer pair for amplifying target sequence, a plasmid group and application thereof. The sgRNA group designed by the present application edits the sheep DYA gene, and after gene editing, large fragment deletion occurs and causes frame shift mutation, which causes great damage to the protein structure and function encoded by the DYA gene. The experimental results show that different fragment lengths of gene editing can be caused in the target region, the overall editing efficiency of the first exon reaches 76.4%, and the overall editing efficiency of the second exon reaches 84.09%. Editing occurs on both sets of chromosomes in the genome (i.e. homozygote): the homozygote editing efficiency of the first exon is 30.9%, and the homozygote editing efficiency of the second exon is 9.1%. The sgRNA group provided by the present application has important significance in the study of ruminant evolution and related immune mechanisms.

A chimeric helper nucleic acid molecule contains polynucleotides encoding helper functions from three distinct viruses for producing recombinant adeno-associated virus (rAAV). The molecule includes adenoviral elements with an E2A gene, an E4 gene, and a VA-RNA polynucleotide; a herpes simplex virus (HSV) polynucleotide encoding a UL12 protein or ICP8 protein; and a human bocavirus (HBoV) polynucleotide encoding a NS2 protein or NP1 protein. These polynucleotides are arranged as individual expression cassettes under non-endogenous promoter control. The chimeric helper molecule, when used in combination with an AAV transfer plasmid and Rep / Cap plasmid, provides enhanced production of infectious rAAV particles compared to conventional helper plasmids containing only adenoviral elements. The molecule enables efficient production of multiple AAV serotypes across different cell lines and transfection conditions.

The invention discloses a construction method and application of an NDST3 knock-down plasmid, and belongs to the technical field of biomedicine. Comprising the following steps: S1, querying a serial number of NDST3, and screening out a target sequence; s2, annealing the forward oligonucleotide and the reverse oligonucleotide to form a double-stranded DNA fragment with a cohesive end, filling a sequence, exposing the cohesive end, and connecting the fragment with a plasmid vector subjected to the same double enzyme digestion linearization to construct a recombinant plasmid; s3, adding the forward oligonucleotide and the reverse oligonucleotide into 10xNEB buffer 2, then adding ddH2O, diluting to 20 mu M, putting into a PCR (Polymerase Chain Reaction) instrument, keeping at high temperature, and slowly cooling to room temperature to obtain ligos with the concentration of 6pmol; s4, carrying out plasmid enzyme digestion treatment, and setting a no-load plasmid group; and S5, adopting Takara-DNA (Deoxyribose Nucleic Acid) Ligalization kit ver.2.1 to connect the oligo and the oligo, and adopting a PCR (Polymerase Chain Reaction) instrument to connect the plasmid, the oligo and 8 [mu] l of solution I at 16 DEG C for 30 minutes. The invention provides transfection plasmids for subsequent WB experiments, and verifies that the expression of the NDST3 is reduced when the shNDST3 plasmids are instantly transferred into human and mouse cells.

The invention relates to the field of biological agents, in particular to application of a plasmid composition to preparation of a medicine for treating bone defect and / or bone remodeling abnormality diseases. The plasmid group composition is a DNA (deoxyribonucleic acid) plasmid composition containing SEQ ID NO: 1 or a DNA plasmid composition containing more than 90% of similarity with the sequence of SEQ ID NO: 1. The plasmid composition provided by the invention has the effect of promoting bone injury repair, can promote bone defect repair and reconstruction, maintains better bone density, and has better healing effect. Meanwhile, the plasmid injection is free of obvious local stimulation and high in safety.

The invention discloses a recombinant bovine rotavirus strain as well as a preparation method and application thereof, and belongs to the technical field of bovine rotavirus vaccines. The invention aims to provide a multi-combined vaccine which takes bovine rotavirus as a carrier and can prevent calf and piglet diarrhea. The invention provides an attenuated bovine rotavirus rescue plasmid combination, which is characterized in that a pT7 vector is used as a starting vector, and the starting vector is respectively connected with a pT7 / VP1BLR plasmid, a pT7 / VP2BLR plasmid, a pT7 / VP3BLR plasmid, a pT7 / VP4BLR plasmid, a pT7 / VP6BLR plasmid, a pT7 / VP7BLR plasmid, a pT7 / NSP1BLR plasmid, a pT7 / NSP2BLR plasmid, a pT7 / NSP3BLR plasmid, a pT7 / NSP4BLR plasmid and a pT7 / NSP5BLR plasmid which are obtained by sequences as shown in SEQ ID NO.2-12. A research and development thought is provided for constructing a multi-combined vaccine.

This invention discloses a replication-deficient recombinant rabies virus chimerically expressing feline herpesvirus type I protein and its applications. It belongs to the field of biomedical technology. The purpose of this invention is to provide a genetically engineered vaccine for feline infectious rhinotracheitis. This invention provides a replication-deficient recombinant rabies virus chimerically expressing feline herpesvirus type I protein, using plasmid combination 1 or plasmid combination 2 to rescue rabies virus strains; plasmid combination 1 is shown below: pD-N, pD-P, pD-G, pD-L, and pD-SRV9-△G-gB; plasmid combination 2 is shown below: pD-N, pD-P, pD-G, pD-L, and pD-SRV9-△G-gD. The goal is to develop a safe and effective novel vaccine.

A plasmid combination for preparing an extracellular vesicle loaded with a target protein or polypeptide, including a pCAG-GOI plasmid and a pMD2.G plasmid, where GOI refers to a gene sequence corresponding to the target protein or the polypeptide. The pCAG-GOI plasmid is a pCAG-cyclic GMP-AMP synthase (cGAS) gene plasmid, a pCAG-cyclization recombinase (Cre) gene plasmid and a combination thereof. An extracellular vesicle loaded with cyclic GMP-AMP synthase, cyclization recombinase or a combination thereof is provided, which is prepared by transfecting a cell with the plasmid combination. An application of the extracellular vesicle in the prevention or treatment of a cancer is further provided.

The application discloses a recombinant bovine rotavirus strain and a preparation method and application thereof, and belongs to the technical field of bovine rotavirus vaccines. In order to provide a multi-vaccine which can prevent diarrhea of calf and piglet and takes bovine rotavirus as a carrier, the application provides a combination of attenuated bovine rotavirus rescue plasmids, wherein the bovine rotavirus rescue plasmid is a pT7 / VP1BLR, pT7 / VP2BLR, pT7 / VP3BLR, pT7 / VP4BLR, pT7 / VP6BLR, pT7 / VP7BLR, pT7 / NSP1BLR, pT7 / NSP2BLR, pT7 / NSP3BLR, pT7 / NSP4BLR and pT7 / NSP5BLR plasmid obtained by connecting sequences shown in SEQ ID NO. 2-12 respectively and taking a pT7 vector as a starting vector. The application provides a research and development idea for constructing a multi-vaccine.

The present invention belongs to the field of microbial genetic engineering technology, and discloses a construction method and application of an Escherichia coli engineered strain with high lactoyl-N-neotetraose production. The Escherichia coli engineered strain with high lactoyl-N-neotetraose production of the present invention overexpresses recombinant genes, including β-1,3-N-acetylglucosamine aminotransferase gene lgtA, UDP glucose 4 epimerase gene galE, and β 1,4 galactosyltransferase gene, thereby increasing LNnT production by means of plasmid combination optimization, protein tag co-expression, related gene knockout, and genome integration. The present invention selects an Escherichia coli engineered strain to increase the production of LNnT in shake flask fermentation from 0.67 g / L to 2.66 g / L, and the highest LNnT production in 5L tank fermentation reaches 17.26 g / L.

The invention belongs to the field of gene engineering, and particularly relates to a non-interval loop iteration yeast genome editing system. The system is composed of Cas and gRNA double plasmids, wherein the gRNA plasmids comprise auxotrophy gRNA-URA3, gRNA-HIS3 and gRNA-LEU2, and the gRNA plasmids comprise gRNA-URA3, gRNA-HIS3 and The resistance is gRNA-BleoR, gRNA-HygR, gRNA-NrsR, gRNA-BleoR, gRNA-HygR and gRNA-NrsR; through coordinated use with a Cas selection marker, the method realizes interval-free, cyclic and iterative efficient editing operation on a yeast genome, and can be used as a practical tool for multi-gene continuous iterative editing of saccharomyces cerevisiae and other fungi. The method has extremely high editing efficiency, an experimental terminal eliminates Cas and gRNA plasmids to realize traceless operation, and the method is low in off-target rate, universal, accurate and easy to operate and can be applied to large-scale gene knockout, knockin, site-specific mutagenesis and the like.

The invention belongs to the field of biological agents, and particularly relates to application of a plasmid composition in preparation of a medicine for promoting healing of a dental wound area. The plasmid group composition is a DNA (deoxyribonucleic acid) plasmid composition containing SEQ ID NO: 1 or a DNA plasmid composition containing more than 90% of similarity with the sequence of SEQ ID NO: 1. The plasmid composition provided by the invention can shorten the repair and reconstruction time of alveolar bone and maintain better bone density, alveolar ridge height and width; the plasmid composition is free of obvious local stimulation, high in safety and good in application prospect.