161 results about "Structural protein" patented technology

The invention discloses a variable region amino acid sequence of a monoclonal antibody of a West Nile virus non-structural protein NS1 and application of the variable region amino acid sequence, and belongs to the technical field of medicines. According to the invention, West Nile virus non-structural protein NS1 expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the West Nile virus non-structural protein NS1 are screened from rabbit spleen cells through flow sorting, and a signal peptide and a variable region gene fragment of an antibody are cloned through reverse transcription-polymerase chain reaction; according to the present invention, the non-structural protein NS1 of flaviviridae flaviviridae virus is taken as a template, and is connected with a constant region gene to an expression vector, and after mammalian cell expression and purification, the monoclonal antibody which has high affinity and is not combined with the non-structural protein NS1 of other eight viruses of flaviviridae flaviviridae virus is obtained through enzyme-linked immunosorbent assay; the monoclonal antibody has application value in diagnosis and prevention and treatment of West Nile virus infection.

This invention provides a dengue / Zika virus genetically engineered vaccine and its application. The dengue / Zika virus vaccine comprises an open reading frame encoding envelope protein domain III (EDIII) and the non-structural protein NS1, and displays EDIII monomers in the delivery vector shell. Immunization with this vaccine primarily induces a type-specific antibody response, reducing the production of cross-antibodies and thus effectively avoiding or eliminating the risk of antibody-dependent enhancement of infection (ADE). This vaccine can be used to prevent dengue virus and Zika virus infection.

The application provides a PDCoV virus mRNA vaccine which can be self-cleaved and expresses multiple virus structure proteins and a preparation method thereof, the vaccine comprises mRNA for expressing PDCoV virus S, M and N proteins, and LNP for encapsulating the mRNA, denoted as SMN-mRNA-LNP. The application firstly proposes a PDCoV mRNA vaccine strategy based on S, M and N combination, connects three structure proteins of PDCoV S, M and N by using a self-cleaving peptide P2A, mutates the S protein with double proline, enables a single mRNA to express multiple PDCoV antigens, and utilizes the neutralizing antibody induction ability of the S protein, the immune regulation function of the M protein and the cell immune activation characteristics of the N protein to construct a multi-level defense system. Through evaluation of immunogenicity, antibody level and challenge protection effect of the vaccine in mice, suckling piglets and pregnant sows, a new idea for developing a broad-spectrum and high-efficiency PDCoV vaccine is provided, and practical basis for research and development of a coronavirus multi-antigen mRNA vaccine is provided.

The invention relates to compositions and methods for the preparation, manufacture and therapeutic use of oncolytic defective virus compositions and methods of in vivo synthesis thereof. The composition includes a first nucleic acid construct encoding a self-amplifying mRNA (sa-mRNA) encoding at least one gene of interest (GOI) or a plurality of GOIs, a second nucleic acid construct encoding an mRNA encoding at least one virus structural protein, and at least one payload delivery system.

Disclosed herein are vaccine constructs for producing a virus-like particle (VLP) capable of raising an immune response to an immunogen, and uses thereof, wherein the constructs comprise nucleic acid sequences encoding an immunogen and a polyprotein, wherein the polyprotein comprises two or more viral structural proteins, wherein at least two of the two or more viral structural proteins are separated by a signal peptidase sequence such that, when the polyprotein is expressed in a host cell, the signal peptidase sequence undergoes host cell peptidase-dependent cleavage to liberate the two or more viral structural proteins, thereby allowing the liberated structural proteins to self-assemble into a VLP carrying the immunogen.

The invention discloses a skin barrier repairing oral liquid based on cubilose peptide-collagen-traditional Chinese medicine compounding. The skin barrier repairing oral liquid is prepared from the following raw materials in percentage: 30 to 50 percent of skin nutrition base powder, 5 to 10 percent of three-type collagen peptide, 0.6 to 1 percent of cubilose peptide and 30 to 70 percent of traditional Chinese medicine herbal extract. The three-type collagen peptide disclosed by the invention can be used for directly supplementing key structural protein of a dermis layer of skin, promoting collagen fiber regeneration, improving skin elasticity and reducing fine wrinkles; the cubilose peptide is small in molecular weight, easy to absorb and capable of providing amino acid raw materials for skin cells and enhancing cell activity, the cubilose peptide and the collagen peptide cooperate to solve the problem of insufficient collagen synthesis caused by age increase, the skin nutrient base powder, the collagen peptide and the cubilose peptide are nutrients needed by skin barriers, and the herbal medicine is used for promoting nutrient absorption, mobilizing qi and blood of the body and improving the taste.

The invention relates to the field of biological medicine, and provides a coxsackie virus A16 type mutant and a virus-like particle thereof, compared with a wild type coxsackie virus A16 type, the mutant has at least one amino acid mutation site in the following regions: (1) a canyon region of a virus capsid; (2) a channel area at a secondary axis; or (3) a channel region at a quasi-tertiary axis. The mutant and the virus-like particle are obtained by optimizing and modifying structural proteins VP0, VP1 and VP3 which form a virus capsid by using computational biology and structural biology methods, co-expressing and co-assembling the structural proteins in a hansenula polymorpha expression system to form the virus-like particle, and then carrying out series of chromatographic purification. Compared with a non-mutated coxsackie virus A16 type virus-like particle mutation scheme, the coxsackie virus A16 type virus-like particle mutation scheme disclosed by the invention has the advantages that the antigen immunogenicity can be obviously improved, and the clinical application value is realized.

A provided self-replicating mRNA is transcribed from an alphavirus backbone vector. The alphavirus backbone vector comprises gene sequences of alphavirus non-structural proteins nsP2, nsP3, nsP4, and mutated nsP1. A cysteine at position 492 of the mutated nsP1 is mutated into serine. The self-replicating mRNA has a higher expression level and a longer expression time for a target gene. The self-replicating mRNA is used for expressing IMP3, and can be prepared into a vaccine having good preventive and therapeutic effects on IMP3-positive tumors. Therefore, the present invention has good application prospects in drug and vaccine development.

A lipid nanoparticle composition for use in the delivery of a self-amplifying RNA (saRNA) construct in one or more target cells is disclosed. The lipid nanoparticle composition comprises a lipid mixture comprising at least one (ionizable) cationic lipid, at least one helper lipid, a sterol, a corticosteroid such as Dexamethasone, and a least one lipid-polyethylene glycol conjugate. The saRNA construct comprises a first open reading frame which encodes one or more non-structural proteins, and a second open reading frame operatively linked to the first open reading frame. The second open reading frame comprises a coding region which encodes one or more target proteins. In some embodiments, the target proteins comprise one or more therapeutic proteins. In some embodiments, the target proteins comprise one or more one or more Cas proteins and / or variants thereof for use in CRISPR-based gene editing.

The present invention discloses a recombinant virus-like particle of Akabane disease virus and a preparation method thereof, and belongs to the field of biotechnology. The recombinant virus-like particle of Akabane disease virus is obtained by recombinantly expressing the structural protein Gn and the structural protein Gc of Akabane disease virus in an insect cell-baculovirus expression system and self-assembly; the amino acid sequence of the structural protein Gn is shown in SEQ ID NO.1; the amino acid sequence of the structural protein Gc is shown in SEQ ID NO.2. The present invention optimizes the genes encoding the structural proteins Gn and Gc according to the codon preference of insect cells, and successfully prepares safer AKAV virus-like particles through the baculovirus-insect cell expression system. The AKAV virus-like particles provided by the present invention have application potential in various fields such as AKAV vaccines and immunotherapy, and provide technical reserves for biosafety monitoring.

The application discloses a preparation method of a decellularized matrix and a product prepared by the method. The preparation method comprises the following steps: (1) washing a mammalian soft tissue to remove blood, lipids and impurities, and cutting the tissue into pieces with a thickness of 1-10 mm; (2) soaking the pieces in solution 1 for 6-48 h, and then soaking the pieces in solution 2 for 0.1-1 h; (3) treating the pieces in an ultrasonic water bath under the condition of a frequency of 20-40 kHz for 5-30 min; (4) water-bathing the pieces at 55-65 DEG C for 10-30 min; (5) sequentially washing the tissue with deionized water, a PBS buffer and ethanol, and washing the tissue with each solution for 1-3 times, each time for 10-30 min; and (6) freeze-drying the decellularized matrix material, and crushing the material at-80 DEG C to-20 DEG C to obtain a decellularized matrix powder. The preparation method significantly improves the cell component removal efficiency, effectively protects structural proteins such as collagen from being degraded in the freeze-drying and crushing processes, and ensures the integrity of the matrix in the aspects of morphology, structure and function, thereby providing a structural support and biological signal support for subsequent tissue repair.

The present disclosure relates to a novel system for the generation and use of hybrid alpha-pseudoviruses against ribose viruses. A hybrid ribose virus-alpha-pseudovirus (HRAP) presents a novel particle having an RNA genome derived from an alphavirus and a structural protein of a virus in the ribose virus field. The HRAP particle is formed by assembling structural proteins crossing different riboviridae and can be used for vaccine development, antiviral drug screening, neutralization test, treatment or immune response starting and the like.

This invention provides the application of apatinib in the preparation of antiviral drugs, specifically involving the application of the anticancer drug apatinib in inhibiting the replication of Hantanensis virus (HTNV) and Chikungunya virus (CHIKV). Apatinib can significantly inhibit HTNV replication and reduce viral titers at the cellular level. Experiments have shown that after treatment with apatinib, the expression levels of viral structural proteins, the nucleic acid levels of HTNV, and the viral titers in the cell culture supernatant were significantly reduced in HTNV-infected A549 cells, while the drug exhibited low cytotoxicity. Furthermore, apatinib can also inhibit CHIKV replication. This invention provides a novel anti-HTNV candidate drug and offers insights for the rapid development of antiviral drugs.

This invention belongs to the field of genetic engineering. By introducing a TC tag between amino acids 199 and 200 of the wild-type bluetongue virus non-structural protein NS2, a recombinant virus was rescued. Analysis of viral plaques and growth curves revealed that the recombinant virus formed plaques of similar size to the wild-type virus, and their growth curves showed no significant difference. Immunofluorescence and FlasH-EDT2 staining of NS2 showed that the fluorescence of NS2 labeled with anti-NS2 antibody overlapped with that labeled with FlasH-EDT2. Three-dimensional imaging of FlasH-EDT2-labeled NS2 revealed that the viral inclusion bodies exhibited fluorescence around the periphery but no fluorescence inside, displaying an overall "core-shell" hierarchical structure. This recombinant bluetongue virus can be used for tracing and localizing NS2 protein and viral inclusion bodies in living cells, and can also be applied to BTV neutralization assays, antiviral drug screening, and other related research.

This invention discloses an anti-phage system based on ESCRT-III and its applications, relating to the field of biotechnology. This invention utilizes the coding genes for two structural proteins of the Hoda_Snf7_1 and Hoda_Snf7_2 from the Hoda archaea ESCRT-III. Hoda_ snf7_1 and Hoda_snf7_2 This invention is applied to antiphage systems. Compared with existing technologies, the main advantages of the antiphage system of this invention are: (1) it consists of only two genes, making system construction simple and quick; (2) it is composed of structural proteins, making it less prone to losing antiphage activity due to gene mutations; (3) it has a broad-spectrum antiphage effect; and (4) it does not affect the normal growth of host bacteria. This system expands the understanding of antiphage systems and provides new ideas and technical means for preventing and controlling phage contamination in industrial microbial production processes.

The invention belongs to the technical field of food processing, and particularly relates to a preparation method and application of a meat anti-freezing adhesive containing plant polyphenol and ice structuring protein. The meat adhesive comprises the following components in percentage by mass: 0.1%-5% of ice structuring protein, 0.5%-10% of plant polyphenol, 9%-14% of glutamine transaminase, 30%-60% of sodium caseinate, 5%-20% of gelatin, 2%-5% of sodium tripolyphosphate, 2%-5% of sodium pyrophosphate and 10%-25% of maltodextrin. The invention provides a natural, efficient and multifunctional meat anti-freezing adhesive which can effectively inhibit ice crystal growth and recrystallization in the freezing process, reduce juice loss, firmly bond meat pieces and delay oxidation discoloration.

The invention discloses a modified phoenix ovi cornea repair material as well as a preparation method and application thereof, an EDTA-imidazole mixed solution is used for efficiently removing a calcium carbonate shell layer of an egg, and no calcium residue exists. Soaking treatment is carried out under the optimized alkali concentration to realize partial denaturation of main structural protein, so that the natural fiber network of the hen egg membrane becomes loose, and the optical clarity is improved while the mechanical integrity is kept. Alkaline treatment further improves the physicochemical characteristics of the membrane, namely surface hydrophilicity, balanced water content and degradation rate, the saturated water content of the modified phoenix membrane is close to that of natural cornea, the degradation rate is matched with the regeneration and repair rate of cornea defects, and the phoenix membrane is endowed with excellent antibacterial performance through quaternary ammonification reaction. A New Zealand white rabbit anterior lamellar cornea cutting operation model is adopted to evaluate the cornea regeneration and repair curative effect, a modified phoenix membrane cornea repair material is implanted, coherent epithelium closure and ordered matrix remodeling of the cornea are effectively promoted, and the repaired cornea is consistent with a natural lossless cornea in function.

The invention provides a norovirus genome splicing method and device, a medium and a product. The method comprises the following steps: acquiring a norovirus gene database, wherein the norovirus gene database comprises a non-structural protein coding region sub-library and a capsid protein coding region sub-library; obtaining original sequencing data; performing quality control on the original sequencing data to obtain screened sequencing data; comparing and assembling the screened sequencing data with the non-structural protein coding region sub-library, and determining at least one first recombinant coding region; and comparing and assembling the screened sequencing data with the capsid protein coding region sub-library, and determining at least one second recombinant coding region, and splicing the at least one first recombinant coding region and the at least one second recombinant coding region to obtain a recombinant genome sequence. Through the technical scheme disclosed by the invention, efficient and accurate splicing of the recombinant norovirus genome sequence is realized, and the assembly accuracy and efficiency of the recombinant norovirus genome sequence are improved.

The invention relates to the technical field of biology, and discloses a Zika virus non-structural protein 1 functional site and application thereof in design of a Zika virus attenuated strain. According to the invention, a new Zika virus virulence site is innovatively found, and a basis is provided for research of Zika virus and research and development work of related drugs. The invention also provides an attenuated mutant strain constructed on the basis of the virulence site, and the attenuated mutant strain has obvious attenuated characteristics and higher genetic stability, and can be used as an alternative strain of a Zika virus attenuated live vaccine.

The invention discloses a method for secretory expression of recombinant mechanical structural protein by using escherichia coli and application, and relates to the field of bioengineering. According to the method, signal peptides are introduced to the N end of recombinant mechanical structural proteins, and the signal peptides, namely OmpA, PelB, PhoA, DsbA and STII, capable of guiding different recombinant mechanical structural proteins to secrete and express are screened from six natural signal peptides from escherichia coli and other species. Meanwhile, the yield of extracellular protein is further improved by combining an amino acid tRNA supplement strategy. According to the method, the recombinant escherichia coli strain capable of efficiently secreting and expressing the mechanical structure protein can be obtained, the downstream purification process is simplified, and the method has important application value.

The application discloses a recombinant FCV antigen and a construction method and application thereof. The construction method of the recombinant FCV antigen comprises the following steps: fusing a T cell epitope coding sequence of a non-structural protein NS7 of FCV and a coding sequence of a SpyTag peptide segment through a coding sequence of a linker, then cloning into a baculovirus transfer vector to obtain a recombinant plasmid, and finally integrating the T cell epitope coding sequence into Bacmid through Tn7 transposition to finally obtain the recombinant FCV antigen. The application selects NS7 as a core immunogen, guides the immune system to produce a high cellular immune response, and thus makes up for the deficiency of an existing vaccine in clearing intracellular viruses; meanwhile, a specific T cell epitope is selected in the sequence of NS7 as an immunogen, which can avoid the immunological escape caused by the variation degree of antigens among different strains and virus antigen drift, and thus provides broader protection.

The present application relates to a kind of preparation methods of deformation-resistant high-viscosity food gel, comprising: pure water, soy protein, TG enzyme proportionally sequentially mixed, using stirrer stirring, heating makes protein and TG enzyme fully crosslinking;Add vegetable oil, emulsify fully;Add potato starch, konjac flour, continue to stir until no lump;Mixture is placed in mold and shaped;Heat curing, put into refrigerator and store, obtain deformation-resistant high-viscosity food gel;Gel is placed in coating device, is smeared between any plant meat tissue, is placed in vacuum bag and is packaged, after secondary maturation shaping, form the multi-level structure of plant meat.The effect of protein and TG enzyme promotes gel formation and gel strength increase, by multi-component synergy, form multi-level, high-strength composite network structure, the obtained food gel is deformation-resistant, plastic, can effectively connect different plant protein tissue, also can improve the texture and taste of plant meat, improve the sensory diversity of plant meat.

Provided herein are stoichiometric Gas Vesicle Expression Systems (GVES), sets of polynucleotide constructs, and related vectors, cells, compositions, and methods configured for robust expression of Gas Vesicle Gene Clusters (GVGCs) in mammalian cells, particularly primary and immune cells. The GVES comprises distinct gene modules for the primary structural protein (gvpA / B) and assembly factors (AF1, AF2), operably configured to achieve a stoichiometric expression ratio, quantified by Dosage Index (DI), wherein the gvpA / B module is expressed at least 2-fold higher than the AF1 and AF2 modules.

The invention provides a GP4 protein antigen epitope peptide of PRRSV (Porcine Reproductive and Respiratory Syndrome Virus) and a monoclonal antibody, and provides a hybridoma cell strain for generating the monoclonal antibody, and the preservation number of the hybridoma cell strain is CCTCC (China Center for Type Culture Collection) NO.C2025259. The monoclonal antibody provided by the invention does not react with various other swine viruses such as PCV2, PEDV, GETV and the like, and has good specificity; the polypeptide has good affinity with PRRSV structural protein GP4 at the molecular level and the cellular level, and epitope information is clear. The antibody can significantly inhibit the infection efficiency of a PRRSV-2 JXwn06 strain in MARC-145 cells and alveolar macrophages, has a good PRRSV in-vitro neutralization effect, can be used for developing a high-specificity PRRSV-2 GP4 detection reagent, and is expected to be used as a candidate drug for passive immune prevention and treatment of PRRS.

The invention belongs to the technical field of animal reproductive biology and cell engineering, and particularly relates to a fluorescence labeling expression system for visualizing an oocyte nucleus maturation process and application of the fluorescence labeling expression system. The fluorescence labeling expression system comprises an eukaryotic expression vector pVenus-LMNB1; the eukaryotic expression vector pVenus-LMNB1 comprises a nucleotide sequence for coding the fusion protein Venus-LMNB1. According to the invention, a nuclear fiber layer protein stably expressed on a nuclear membrane is selected as a research object, a eukaryotic expression vector pVenus-LMNB1 is constructed by using a molecular cloning technology, an LMNB1 gene containing a green fluorescent label Venus is overexpressed in a porcine oocyte by using a microinjection technology, fluorescent protein labeling of a structural protein on the nuclear membrane is utilized, and the expression vector pVenus-LMNB1 is constructed. The expression positioning condition of the LMNB1 in the maturation process of the porcine oocytes is observed, and a powerful tool is provided for development of in-vitro culture related researches of the porcine oocytes.

The present invention is in the field of Nipah virus (NiV) structural protein. Particularly, the invention provides structure based stabilized Nipah virus (NiV) structural protein designing and uses thereof.

The present application relates to the technical field of probiotic microcapsules, and particularly relates to a high-activity composite wall material probiotic microcapsule and a preparation method thereof.The high-activity composite wall material probiotic microcapsule comprises a composite wall material and a core material, and the raw material of the composite wall material comprises konjac glucomannan, soybean protein isolate and nano titanium dioxide, and the mass ratio of the konjac glucomannan, the soybean protein isolate and the nano titanium dioxide is (3-5):(2-4):(0.5-1.5).By constructing the composite wall material system composed of the konjac glucomannan, the soybean protein isolate and the nano titanium dioxide, the polysaccharide skeleton structure, the protein flexible network and the nano particle function enhancement effect are mutually synergistic, so that the mechanical properties, the structural stability and the environmental tolerance of the microcapsule are significantly improved.Meanwhile, the nano titanium dioxide endows the microcapsule with excellent antibacterial properties and ultraviolet shielding capacity, and can effectively inhibit the influence of external harmful microorganisms and reduce the damage of light to the activity of the probiotic.

The invention belongs to the field of gene engineering, and particularly relates to application of chikungunya virus structural protein in improvement of pseudovirus stability, a construction method of the chikungunya virus structural protein and vaccines, antibody evaluation and in-vivo gene therapy products. The method comprises the following steps: constructing a human codon optimized Asian strain CHIKV-E3 + E2 + 6K + E1 structural protein eukaryotic vector, and designing two types of binder expression plasmids of targeted T cells; the vector, a psPAX2 helper plasmid and a lentivirus target vector containing an SFFV promoter and a ZsGreen reporter gene (or a CAR therapeutic gene) are co-transformed into a 293T cell, and the lentivirus is prepared through culture, filtration and centrifugation. The lentivirus can efficiently infect Jurkat cells and activate human T cells, obviously reduces the infection efficiency on 293T cells so as to reduce the off-target effect, has excellent stability in human serum, and provides a safe and efficient gene delivery tool for in-vivo CAR-T therapy.

The invention provides an application and a method of GSH in promoting mature differentiation of hiPSC-CMs, and the method is characterized in that in the process of directionally differentiating the hiPSC-CMs into the hiPSC-CMs, an effective dose of GSH is added into a differential medium, and the optimal concentration is 20-100 [mu] M. According to the invention, through simple and economical exogenous supplement of GSH, maturation of hiPSC-CMs is systematically promoted from multiple dimensions, including enhancement of intracellular anti-oxidation defense capability; energy metabolism is promoted to be converted from glycolysis to oxidative phosphorylation; up-regulating expression of cardiac muscle cell specific structural protein and calcium ion channel related genes; mitochondrial biological generation and functions are promoted; and the cell morphological structure is induced to develop to a mature phenotype. The method is simple and convenient to operate and high in biocompatibility, the hiPSC-CMs with mature functions can be stably obtained, and a high-quality cell source is provided for application of the hiPSC-CMs in the fields of heart disease modeling, drug cardiotoxicity screening and the like.

Synthetic peptides mimicking the nsp10 sequence in the region interacting with nsp16 capable of penetrating cell membranes and inhibiting SARS-CoV-2 replication for the treatment of moderate to severe COVID-19. The invention relates to peptides inhibiting SARS-CoV-2 replication, likely through inhibition of Methyltransferase complexes (NSP10 / NSP16 and NSP10 / NSP14). The peptide of the present invention, P3, contains sequences corresponding to amino acids 89-96 of the non-structural protein 10 (NSP10) of SARS-CoV-2, with the only Cysteine modified to a Methionine. This peptide was made based on two previous designs P1 and P2, which constituted the amino acids 68-96 of the NSP10 protein of SARS-CoV-2.