23results about How to "High infection efficiency" patented technology

The invention provides a recombinant HSV-1 vector, oncolytic HSV-1 and application of the oncolytic HSV-1, and belongs to the technical field of viral vectors. According to the recombinant HSV-1 vector provided by the invention, nucleotides at the 117014 site to the 132565 site and nucleotides at the 145316 site to the 145585 site in an HSV-1 genome are deleted. The recombinant HSV-1 vector provided by the invention is high in safety, can be copied in mammalian cells and carry a plurality of exogenous genes, has the advantages of wide host range, high infection efficiency, large capacity, easiness in preparation and sustainable expression of the carried exogenous genes, and can be applied to the fields of tumor treatment, gene therapy and virus vector vaccines. The oncolytic HSV-1 provided by the invention is high in safety, retains the replication ability in mammalian cells, has a remarkable oncolytic effect, and can induce an organism to generate tumor immune response so as to play a tumor inhibition role.

The invention discloses a yak intramuscular precursor fat immortalized cell line and a preparation method thereof. The method comprises the following steps: (1) obtaining yak primary intramuscular precursor fat cells; (2) performing SV40T lentivirus infection on the precursor fat cells in the yak muscle and screening cell strains: performing SV40T lentivirus infection by utilizing a 1 / 2 small-volume infection method to obtain immortalized cells after the precursor fat cells in the yak primary muscle are infected with SV40T, namely immortalized cells for short; puromycin is adopted to screen immortalized cells; and diluting the screened cells by using a limited dilution method, and carrying out enlarged culture to obtain the immortalized cell line. The yak intramuscular precursor fat immortalized cell line constructed by the invention has genetic homogeneity, the heterogeneity influence of cell populations is eliminated, and an in-vitro cell model is provided for subsequent research related to yak intramuscular fat cells.

The present application relates to the field of biomedicine, and specifically to a fusion adeno-associated virus capsid protein, adeno-associated virus and its application. The present application provides a fusion adeno-associated virus AAV-WM14 capsid protein, comprising amino acid fragments of serotypes AAV1, AAV2, AAV3, AAV4, AAV6, AAV7, AAV8, AAV10, and AAV13, and the capsid protein includes the amino acid sequence shown in SEQ ID NO.1. The fusion adeno-associated virus provided in the present application can be administered by intravitreal injection, efficiently infecting retinal photoreceptor cells in the eye, and can be delivered to the cochlea to efficiently infect inner hair cells and supporting cells. The fusion adeno-associated virus provided in the present application can be widely used in gene therapy for related diseases.

The invention belongs to the technical field of biological medicine, and particularly relates to a method for improving immune cell virus infection efficiency and application. The method provided by the invention comprises the following steps: S01, activating immune cells; s02, adding the activated immune cells into a freezing medium; s03, the immune cells added with the cryopreservation liquid are subjected to rapid cryopreservation under the condition of-80 DEG C to-70 DEG C; and S04, quickly thawing the frozen immune cells in a water bath at 35-37 DEG C, wherein the temperature of the water bath is 35-37 DEG C. According to the method provided by the invention, the immune cells with improved virus infection efficiency can be obtained, in addition, the method does not destroy the normal killing function of the immune cells, and the virus infection positive rate of the treated immune cells is more lasting.

The application provides an AAV9 capsid protein variant and use thereof. Research finds that the AAV9 capsid protein is modified, and the modified amino acid sequence is shown in SEQ ID NO: 1, which can significantly escape neutralizing antibodies, and has significant positive significance for gene therapy.

This invention discloses a recombinant oncolytic virus based on a Luyo virus G protein mutation and its large-scale production method. The method involves using reverse genetics to replace the G protein-coding gene of a wild-type oncolytic vector with a mutated Luyo virus G protein-coding gene, while simultaneously inserting a functional cytokine-coding gene into the wild-type oncolytic vector, thereby rescuing and obtaining a self-replicating recombinant oncolytic virus. The sequence of the mutated Luyo virus G protein-coding gene is shown in SEQ ID No. 1. This invention retains ideal therapeutic properties and achieves efficient amplification in interferon signal-deficient cells, facilitating titer determination.

The invention discloses a lentivirus titer detection method using suspension cells. The method comprises the following steps: 1) paving a proper amount of suspension growth cells with determined cell quantity in a 24-pore plate; 2) planking the suspension growth cells, respectively adding different gradients of to-be-detected titers of lentiviruses and lentivirus infection enhancing liquid, and incubating for 5 days under the conditions of 37 DEG C and 5% CO2 to obtain suspension tool cells infected by the lentiviruses; and (3) measuring the tool cells infected by the lentivirus by using flow cytometry or quantitative PCR (Polymerase Chain Reaction) to calculate the titer. The method comprises the following steps: selecting specific suspension growth cells such as Jurkat, Raji, NALM-6 and K562 as tool cells; meanwhile, two different lentivirus enhanced infection solutions A (HitransG A) and P (HitransG P) are combined, WPRE, gag and the like are selected as LVs characteristic genes, a human albumin gene is selected as a host cell characteristic single copy gene and is used as a qPCR detection reference gene for lentivirus titer evaluation, and a set of lentivirus titer detection method which is rapid, accurate, low in cost and repeatable is constructed.

The invention discloses application of a soybean histone variant to improvement of genetic transformation efficiency of soybeans and other plants. By applying the soybean histone variant, the infection efficiency of agrobacterium to soybean cotyledonary node explants can be remarkably improved, so that efficient introduction of exogenous genes is promoted, the transformation efficiency is remarkably improved, finally, the higher stable transformation rate is obtained, and the application can be expanded to recalcitrant plant species with genetic transformation difficulty such as tea. In addition, the subsequent genetic transformation efficiency can also be remarkably improved by using the genetic transformation material for stable overexpression of the soybean histone variant as a receptor.

The invention belongs to the technical field of microorganisms, and particularly relates to a cordyceps sinensis strain with a high infection rate and application of the cordyceps sinensis strain, the classification name of the cordyceps sinensis strain KK1 is Hirsutella sinensis, the cordyceps sinensis strain KK1 is preserved in China General Microbiological Culture Collection Center on June 7, 2024, the address is No. 3, No.1 Yard, Beichen West Road, Chaoyang District, Beijing, and the preservation number is CGMCC NO.969. The preservation number of the strain is CGMCC (China General Microbiological Culture Collection Center) NO.41330, and the ITS sequence of the strain is shown as SEQ ID NO.1. The strain is applied to artificial cultivation of cordyceps sinensis and has high infectivity; the composition can also be applied to skin care products with a skin photoaging repairing function, is high in antioxidant and anti-inflammatory activity, and can effectively improve skin symptoms related to photoaging. The method provides a new technical support for artificial culture of cordyceps sinensis and prevention and treatment of skin photoaging, and has a good application prospect.

Provided are a chimeric oncolytic adenovirus simultaneously expressing IL-12, IFN-γ, and CCL5, and an application thereof in tumor treatment. A capsid protein hexon of the oncolytic adenovirus is formed from chimerizing hexon sequences of the two serotype viruses Ad5 and Ad48, and a fiber protein is formed from chimerizing fiber sequences of the two serotype viruses Ad5 and Ad11. The chimeric oncolytic adenovirus can activate intrinsic anti-cancer activity of a variety of viral structural proteins, the ability to infect tumor cells is increased while also ensuring that the virus itself avoids interception from pre-existing neutralizing antibodies and adhesion and uptake of hepatocytes, and an effect of killing cancer cells is enhanced.

The invention relates to an anti-Theras gene derived from a cold spring, a screening method, an activity determination method and application of the anti-Theras gene, and particularly discloses an anti-Theras gene derived from the cold spring, and the nucleotide sequence of the anti-Theras gene is shown as SEQ ID NO: 1. According to the application, a novel anti-Theras gene is identified from a deep sea cold spring environment for the first time, and heterologous expression and plaque experiments prove that the novel anti-Theras gene can effectively antagonize a Theras defense system of a host. According to the anti-Theras gene provided by the invention, the types and sources of anti-defense genes are expanded, and a gene blank for non-classical defense system Theras from a cold spring environment is provided.

The invention provides an AAV9 capsid protein variant and application thereof. Researches find that the AAV9 capsid protein is modified, the modified amino acid sequence is shown as SEQ ID NO: 1, and the AAV9 capsid protein can significantly escape a neutralizing antibody and has significant positive significance for gene therapy.

The invention belongs to the field of biotechnology, viral vectors and cell therapy, and particularly relates to a preparation method and application of engineered immune cells and adenoviruses. An Ad35 fiber region of an Ad5F35 sequence is replaced by an Ad50 fiber region to construct the Ad5F50 type adenovirus, the Ad5F50 type adenovirus is used for infecting immune cells, especially macrophages, so that the infection efficiency is improved, the infection time is shortened, the macrophages are modified through a chimeric receptor CAR, the phagocytic function of the macrophages is redirected, and the targeted anti-tumor treatment effect is improved.

This invention discloses a viral vector targeting KIF4A to inhibit angiogenesis, its preparation method, and its applications. The viral vector contains three shRNA gene fragments targeting the KIF4 protein-coding region, as shown in SEQ ID NO.1-SEQ ID NO.3. Using a lentiviral packaging system, a stable KIF4A knockdown cell line can be constructed, effectively inhibiting endothelial cell proliferation and angiogenesis, and also inhibiting the paracrine promotion of angiogenesis by glioma cells, providing a new strategy for selecting targets for anti-angiogenic tumor therapy.

The application provides an AAV9 capsid protein variant and use thereof. The inventors have found, through research, that by modifying the AAV9 capsid protein, changing the amino acid sequence of the capsid protein to a sequence as shown in SEQ ID NO: 3, the infection capacity can be significantly improved, especially the transduction efficiency in the liver, thereby providing a new prospect for gene therapy.

This invention belongs to the field of agricultural microbiology technology, specifically relating to *Pseudomonas vensuelae* (…). Pseudocitrobacter vendiensis This invention relates to Pv-LC32 and its application in enhancing the pathogenicity of *Metarhizium anisopliae* against locusts. For the first time, this invention isolates a strain of *Pseudomonas vensuelae* from the midgut of locusts, exhibiting conditional synergistic pathogenicity. This strain does not directly kill the host, but significantly enhances fungal infection efficiency through interaction with *Metarhizium anisopliae*. This discovery provides a new perspective on understanding the interaction mechanism between insect gut symbiotic bacteria and pathogenic fungi, and lays a theoretical foundation for developing biocontrol enhancement technologies.

The application provides a composition, a method and an application for improving the titer of mouse leukemia virus, and belongs to the technical field of biotechnology.The method comprises the following steps: before a host cell is infected with xenotropic mouse leukemia virus, the host cell is pretreated by using SC79; during virus infection of the pretreated host cell and during cell passage, mifepristone is added to a culture medium; and during continuous virus harvesting, LR3 IGF-1 is added to the culture medium.Through adding additives with different functional mechanisms at different stages of virus production, the application realizes dynamic optimization of the whole process of virus production.The experimental results show that the combination of SC79, mifepristone and LR3 IGF-1 can increase the titer of xenotropic mouse leukemia virus by 14.32 times, which is significantly better than the effect of single additive or two additives in combination.The application solves the problems of low titer of virus stock solution and low production efficiency in the prior art.

The present invention discloses a method for constructing CAR-T targeting liver cancer-related antigen DLK1. The present invention selects a single-chain antibody of liver cancer-related antigen DLK1 to be recombined in T cells, and the T cells after genetic engineering are further activated, thereby enhancing the killing power of T cells against liver cancer cells; the single-chain antibody of DLK1 is obtained by obtaining RNA from a hybridoma cell line that secretes DLK1 monoclonal antibody, and obtaining cDNA after reverse transcription; designing PCR primers for heavy chain variable region and light chain variable region; using the cDNA as a template, amplifying and splicing the heavy chain and light chain variable regions to obtain. The detection method of the present invention has relative sensitivity and specificity, and can reduce the side effects of T cell recognition of normal cells. The present invention provides a certain research basis for CAR-T cell immunotherapy in liver cancer immunotherapy.

The invention discloses a transcription factor gene for promoting adventitious root generation of apple dwarf rootstock and a regulation and control method, and belongs to the technical field of plant genetic engineering. The method comprises the following specific regulation and control operations: cloning a transcription factor gene shown as SEQ ID No.1 in a nucleotide sequence table to an overexpression vector in an apple dwarfing rootstock with difficulty in adventitious root generation; transferring agrobacterium tumefaciens EHA105 by using a freeze thawing method to obtain strain cells, and culturing in a culture medium; centrifugally collecting thalli, and suspending with an infection solution to obtain an infection buffer solution; immersing the apple dwarf rootstock tissue culture seedlings in the infection buffer solution, and carrying out vacuum treatment; and culturing in a cuttage culture medium, and obtaining transplanted seedlings when the length of adventitious roots is greater than 10mm and the rooting number is greater than 3 and reaches the transplanting standard. The instant infection and sugar-free culture technology provided by the invention can quickly promote the occurrence of the dwarf rootstock adventitious roots without a tissue culture technology, and has important economic and social benefits for improving the occurrence of the apple dwarf rootstock adventitious roots.

The present application belongs to the technical field of crop breeding, and relates to a potato breeding method for editing ALS gene. The method comprises the following steps: (1) using agrobacterium transformed with a gene editing tool to infect potato explants, wherein the infection medium used contains CaCl2 with a total concentration of 9-15 mM, and the gene editing tool targets potato ALS gene for editing mutation of ALS gene to make potato resistant to imidazolinone herbicides; (2) after co-cultivation and recovery culture, the potato explants are sequentially subjected to resistant callus induction, resistant bud induction and seedling culture on resistant callus induction medium, resistant bud induction medium and seedling medium to obtain regenerated seedlings, and the resistant callus induction medium, resistant bud induction medium and seedling medium all contain imidazolinone herbicides. By using the method of the present application, potato varieties with ALS gene mutation can be obtained with high infection efficiency and high gene editing efficiency.

The invention provides a construction method and application of conditionally replicative Ad5 / 35 chimeric oncolytic adenovirus, and belongs to the technical field of genetic engineering. The construction method comprises the following steps: carrying out enzyme digestion on a PoAd5 plasmid by endonuclease AsiSI; the method comprises the following steps: carrying out in-vitro gene synthesis on a chimeric Ad5 / 35-Hexon (HVR1 + 5) gene segment, carrying out homologous recombination on the chimeric Ad5 / 35-Hexon (HVR1 + 5) gene segment and linear PoAd5 co-transfected BJ5183 to obtain PoAd5 / 35-H, carrying out ClaI enzyme digestion, carrying out in-vitro gene synthesis on Ad5 / 35-Fiber, and carrying out homologous recombination on the Ad5 / 35-Fiber and linear PoAd5 / 35-H co-transfected BJ5183 to obtain a final plasmid oAd5 / 35-HF; and carrying out PacI enzyme digestion, purifying, and transfecting an HEK293 cell to obtain the conditionally replicative Ad5 / 35 chimeric oncolytic adenovirus. The invention provides an innovative adenovirus vector construction scheme for enhancing the infection ability and the treatment effect of viruses.

The invention belongs to the technical field of microorganisms, and discloses an NTase protein, a screening method thereof and application of the NTase protein in preparation of an anti-phage signal system antagonist and / or an antibacterial preparation. The NTase protein provided by the invention specifically comprises an amino acid fragment with a sequence as shown in SEQ ID NO: 1 or a variant fragment with at least 70% sequence identity with the sequence as shown in SEQ ID NO: 1, the NTase protein has an excellent antagonistic effect on a CBASS system of prokaryotes, and the infection efficiency of viruses on prokaryotic cells can be improved by inhibiting the CBASS system; therefore, a better antibacterial effect is achieved, and the compound has a very excellent application prospect in research and development of novel antibacterial drugs.

The invention relates to the technical field of cell culture, and discloses a tracheal epithelial cell and application thereof, the cell is a guinea pig tracheal epithelial immortalized cell, and the preservation number is CGMCC No. 46763. In the invention, the cell has good susceptibility to respiratory pathogens, and has excellent transfection efficiency. The cells can be used for separating or culturing respiratory pathogens, preparing respiratory pathogen vaccines, screening and / or preparing medicines for preventing and / or treating diseases caused by the respiratory pathogens, serving as host cells for researching the respiratory pathogens, detecting the respiratory pathogens for a non-diagnostic purpose and constructing organoids. The method has high scientific research value and practical application potential.