29 results about "Coat protein" patented technology

A spore of a bacillus subtilis B.subtilis WB600 strain is used as an antigen delivery carrier, spore coat protein C (CotC) is used as anchoring protein, a key protein ATPase gene and a CotC gene of micropterus salmoides iridovirus are fused through a gene fusion technology, a recombinant spore surface display system is constructed, the ATPase antigen is stably displayed on the spore surface, and an oral vaccine is prepared. After being orally applied to fishes, the vaccine can tolerate the gastrointestinal tract environment and is accurately delivered to intestinal related lymphatic tissues to stimulate fish immunity, and finally prevention and control of the iridovirus of the micropterus salmoides are achieved.

The invention discloses a multiplex fluorescent quantitative PCR (polymerase chain reaction) detection kit for prawn culture pathogenic bacteria, which is characterized in that the kit contains primers and probes for detecting the prawn culture pathogenic bacteria, wherein the primers and the probes are respectively used for detecting TDH-related hemolsin genes of vibrio parahaemolyticus, ORF107 genes of white spot syndrome viruses, small subunit ribosomal genes of shrimp enterocytozoon hepatopenaei, 37 kDa coat protein genes of infectious subcutaneous and hematopoietic necrosis viruses, DNA (deoxyribonucleic acid) primer genes of full-eye iridovirus 1 and PirA [pi] genes of pathogens of bacterial prawns acute hepatopancreas necrosis; the method has the advantage that the detection of eight prawn culture pathogens can be realized at the same time. The method comprises the following steps of: obtaining a tcdB gene of a high-pathogenicity vibrio and a hemolsin gene of a vibrio harveyi;

The invention belongs to the technical field of viral nucleic acid molecular diagnosis, and particularly relates to a kit for detecting cucumber green mottle mosaic virus based on enzyme-mediated dual-amplification nucleic acid amplification, a detection method and application. The detection system comprises a primer pair DNA upstream primer (SEQ ID NO.2) and a DNA downstream primer (SEQ ID NO.3) for amplifying a coat protein gene of the cucumber green mottle mosaic virus, and a probe (RNA primer (SEQ ID NO.1)) for detecting a CP gene, a nucleic acid amplification group enzyme system, a signal amplification group enzyme system, a nucleic acid scavenging enzyme, an activating enzyme NTP, a buffer solution and a freeze-drying protective agent, wherein the system is prepared into pre-subpackaged freeze-dried balls. The kit provided by the invention can realize RNA detection of aM concentration, and can realize early screening of cucumber green mottle mosaic virus due to high detection sensitivity.

The base editor systems (“MOBE”) that are derived from the CRISPR / Cas9 protein that enable to simultaneously introduce C⋅G to T⋅A and A⋅T to G⋅C point mutations at distinct genomic loci in living cells, with high efficiency and precision. In the MOBE disclosed herein, a piece of RNA (“the gRNA”) of the CRISPR / Cas9 protein is fused to the deaminase enzymes via a coat protein-aptamer interaction. A reporter plasmid comprising the MOBE system that allows for enrichment of cells with co-occurring orthogonal edits and increased editing efficiency.

This invention belongs to the field of biology, specifically relating to a method for preparing polyclonal antibodies based on a specific fragment of the capsid protein of Chrysanthemum B virus. This invention discloses the antigenic determinant CVB-CP-B and its acquisition method, as well as the antigen CVB-CP-B obtained from the antigenic determinant, and Ab-CVB-CP-B, which facilitates the preparation of antibodies from antigen CVB-CP-B. This invention also discloses the application of the above-mentioned antibodies in the detection of Chrysanthemum B virus.

The invention relates to the technical field of biology, and aims to provide an M13 recombinant bacteriophage with PRRSV broad-spectrum neutralizing activity for displaying NPC and application of the M13 recombinant bacteriophage. A nano antibody-CD163 peptide fragment conjugate NPC shown as SEQ ID NO: 1 is displayed on the surface of the N end of capsid protein III of the M13 recombinant phage, 1-3 NPC molecules are contained on the surface of each phage particle, and the phage is used for neutralizing PRRSV (Porcine Reproductive and Respiratory Syndrome Virus). The recombinant bacteriophage provided by the invention is easy to prepare, extremely high in virus titer and excellent in thermal and pH stability; in Marc-145 and PAMs cells, the compound shows strong neutralizing activity on PRRSV II type 1, 3, 5 and 8 pedigree, so that the compound has broad-spectrum neutralizing capacity; the compound can be used as a PRRSV prevention and control preparation, and a new thought is provided for development of safe and efficient novel antiviral drugs or biological preparations.

The present disclosure relates to single-chain antibodies that specifically bind to the cotton leafroll dwarf virus (CLRDV), and methods for using such antibodies in the serological or molecular detection of CLRDV in plant or aphid samples. In particular, the disclosure relates to single-chain antibodies that recognize the CLRDV coat protein (CP) or the CLRDV n-terminal read-through domain (NRTD).

The invention belongs to the field of biology, and discloses a coating protein for diagnosis of an African swine fever virus antibody, application and a kit, and the coating protein comprises a p22 protein and an E199L protein; the amino acid sequence of the p22 protein is as shown in SEQ ID NO: 1, and the amino acid sequence of the E199L protein is as shown in SEQ ID NO: 3. The coating protein is composed of recombinant proteins of p22 and E199L, the two recombinant proteins can react with ASF positive serum and have good reactogenicity, and the detection kit based on the coating protein is high in detection precision, strong in specificity and good in repeatability.

A modified vector of adenovirus-associated virus serotype 8 (AAV-8) for gene targeting and expression is provided, wherein the modified vector includes serotype coat amino acid sequence, insertion site and insertion amino acid sequence. A 10-amino acid sequence set forth in SEQ ID NO.3 is inserted between amino acids at positions 590 and 591 that are set forth in SEQ ID NO.2 of the AAV-8 serotype coat protein: LARGDSTKSA, wherein amino acids at positions 1, 2 and 10 are protective amino acids, and amino acids at positions 3 to 9 are screened amino acid sequences. In addition, the present invention also discloses construction method and application of the modified vector are also provided.

The application belongs to the technical field of virus nucleic acid molecule diagnosis, and particularly relates to a kit for detecting cucumber green mottle mosaic virus based on enzyme-mediated double amplification of nucleic acid, a detection method and application. The detection system of the application comprises a primer pair for amplifying the coat protein gene of the cucumber green mottle mosaic virus, i.e. a DNA upstream primer (SEQ ID NO. 2) and a DNA downstream primer (SEQ ID NO. 3), a probe for detecting the CP gene (an RNA primer (SEQ ID NO. 1)), a nucleic acid amplification enzyme system, a signal amplification enzyme system, a nucleic acid clearing enzyme, an activating enzyme NTP, a buffer, and a freeze-drying protective agent, which are prepared into a pre-dispensed freeze-dried ball. The kit can realize RNA detection at aM concentration, and early screening of the cucumber green mottle mosaic virus can be realized due to high detection sensitivity.

The application discloses a hybridoma cell strain secreting monoclonal antibody PVY-5, the monoclonal antibody and application, the hybridoma cell strain 4B4D8C3 can secrete anti-potato Y virus monoclonal antibody, the preservation number of the hybridoma cell strain is CCTCC NO: C2022282, the monoclonal antibody PVY-5 has specific immunoreaction with the coat protein of potato Y virus, the heavy chain variable region amino acid sequence of the monoclonal antibody PVY-5 is as shown in SEQ ID NO.6, the light chain variable region amino acid sequence is as shown in SEQ ID NO.7, the sensitivity reaches 1:5120 times dilution by using direct ELISA method detection to the leaf of potato Y virus disease infection, the monoclonal antibody PVY-5 can also be used to prepare ELISA kit, or colloidal gold kit, or nano analog enzyme kit, and the potato Y virus disease can be detected quickly, sensitively, economically and accurately.

The embodiment of the invention discloses an A beta vaccine for preventing and treating Alzheimer's disease and a preparation method thereof. The A beta vaccine comprises a phage vaccine and a recombinant gene vaccine; wherein the bacteriophage vaccine comprises a YXL1 vaccine obtained by integrating a gene sequence as shown in SEQ ID NO: 001 into a pCOMB3X bacteriophage coat protein PIII as a target gene, and a YXL2 vaccine obtained by integrating a gene sequence as shown in SEQ ID NO: 002 into the pCOMB3X bacteriophage coat protein PIII as a target gene; the recombinant gene vaccine comprises a pcDNA A [beta] 3-10 vaccine obtained by integrating a gene sequence as shown in SEQ ID NO: 003 into a pcDNA vector as a target gene, and a pcDNA A [beta] 25-35 vaccine obtained by integrating a gene sequence as shown in SEQ ID NO: 004 into the pcDNA vector as a target gene.

Embodiments of the present disclosure pertain to methods of selecting cyclic peptides that bind to a target by transforming a phage display library with a plurality of nucleic acids into bacterial host cells, where the nucleic acids include phage coat protein genes with a combinatorial region that encodes at least one cysteine and at least one non-canonical amino acid. The transformation results in the production of phage particles with phage coat proteins where the cysteine and the non-canonical amino acid couple to one another to form a cyclic peptide library. Phage particles are then screened against the desired target to select bound cyclic peptides. Amino acid sequences of the selected cyclic peptides are then identified. Additional embodiments pertain to methods of constructing a phage display library that encodes the cyclic peptides. Further embodiments of the present disclosure pertain to the produced cyclic peptides, phage display libraries and phage particles.

The invention provides a plasmid vector and application thereof in nucleic acid detection of tomato brown crinkled fruit virus, the plasmid vector comprises a partial motion protein gene segment, a coat protein gene segment and a partial 3'non-coding region segment of a specific sequence of the tomato brown crinkled fruit virus, the partial motion protein gene segment is as shown in the 1st-549th site of SEQ ID No.1, and the coat protein gene segment is as shown in the 349th site of SEQ ID No.1. The coat protein gene segment is as shown in the 550th to 1031th sites of SEQ ID No.1, and the partial 3'non-coding region segment is as shown in the 1032th to 1182th sites of SEQ ID No.1. As a detection standard substance, the plasmid vector provided by the invention has the advantages of good molecular quality, purity meeting requirements, capability of meeting large-batch indexes, good uniformity and strong stability, target fragments can be detected by various molecular detection methods, and a universal positive control sample and reference are provided for the detection and identification work of the tomato brown crinkled fruit virus.

The invention discloses a real-time fluorescent quantitative PCR (Polymerase Chain Reaction) method for detecting capsicum vein mottle virus in tobacco, which comprises the following steps of: firstly, designing and synthesizing a specific primer pair based on a coat protein CP gene conserved sequence of capsicum vein mottle virus isolate in an NCBI (National Center of Biotechnology Information) database; secondly, extracting total RNA of a tobacco sample infected with the pepper vein mottle virus, and performing reverse transcription to obtain cDNA; carrying out real-time fluorescent quantitative PCR (Polymerase Chain Reaction) amplification by using the obtained cDNA as a template and adopting the specific primer pair; and finally, judging whether the sample carries the target virus or not according to the amplification curve, the melting curve and the Ct value, and realizing quantitative analysis of the virus load through the standard curve. The method is high in specificity and sensitivity, compared with a conventional PCR method, the sensitivity is improved by 10 times, and rapid and accurate detection of the pepper vein mottle virus in the tobacco can be achieved.

The present invention relates to a modified virus-like particle (VLP) of cucumber mosaic virus (CMV) comprising at least one chimeric CMV polypeptide, wherein said at least one chimeric CMV polypeptide comprises, preferably consists of (i) a CMV polypeptide, wherein said CMV polypeptide comprises a coat protein of CMV or an amino acid sequence having a sequence identity of at least 75% with SEQ ID NO:48; and (ii) a polypeptide comprising, preferably consisting of, a stretch of consecutive negative amino acids, wherein said negative amino acids are independently selected from aspartic acid or glutamic acid, wherein said polypeptide is inserted between any amino acid residue of said CMV polypeptide corresponding to any amino acid residue between position 75 and position 85 of SEQ ID NO:48, as well as to compositions and pharmaceutical compositions comprising such modified VLPs to which antigens are linked, which compositions preferably serve as vaccine platform for generating immune responses, in particular antibody responses, against said antigens linked to the modified CMV VLPs.

The application belongs to the field of biology, and discloses a coated protein, application and kit for diagnosing African swine fever virus antibody, wherein the coated protein comprises p22 protein and E199L protein; the amino acid sequence of the p22 protein is shown as SEQ ID NO:1, and the amino acid sequence of the E199L protein is shown as SEQ ID NO:3. The coated protein is composed of recombinant proteins of p22 and E199L, and both of the two kinds of recombinant proteins can react with ASF positive serum, and both have good reactivity. The detection kit based on the above coated protein has high detection precision, strong specificity and good repeatability.

The invention provides a primer for detecting mulberry vein banding virus as well as application and a method. The method comprises the following steps: constructing a standing plasmid system of an MVBaV coat protein N gene, preparing a standard product system, and constructing a standard curve of the standard product system after RT-qPCR (Reverse Transcription-Quantitative Polymerase Chain Reaction). According to the method disclosed by the invention, the detection sensitivity and accuracy of the MVBaV are remarkably improved, the sensitivity is 100 times that of a traditional RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection method, and an efficient and reliable technical means is provided for rapid, qualitative and quantitative detection of the MVBaV.

The application provides a fish bivalent oral vaccine, a preparation method and application, a recombinant plasmid containing dominant antigen epitope sequences of type I and type II grass carp reovirus is electrically transformed into lactobacillus casei to prepare the fish bivalent oral vaccine, and the dominant antigen epitope sequences are shown in SEQ ID NO. 1. The bivalent oral vaccine uses the dominant antigen epitope screened from type I and type II GCRV shell proteins as an immunogen, has special immunogenicity to type I and type II GCRV, the screened dominant antigen epitope sequence encodes a small protein, and the small protein does not cause toxicity and harm to a fish body. After the fish bivalent oral vaccine enters the fish body, the fish bivalent oral vaccine can be expressed in the body and can start an immune response, and can better protect the grass carp from GCRV virus infection and improve the survival rate.

The invention provides a neutralizing epitope polypeptide of grouper nervous necrosis virus and application of the neutralizing epitope polypeptide in serological diagnosis test paper, and the amino acid sequence of the neutralizing epitope polypeptide is SEQ ID NO: 1 or SEQ ID NO: 2. On the other hand, the invention also provides a serological diagnosis test strip for the nervous necrosis virus disease of the grouper, wherein the NNV capsid protein neutralizing epitope polypeptide is used as a detection line. The antigen polypeptide used by the test strip is an NNV capsid protein neutralizing epitope specific fragment, the antigen recognition effect is improved by optimizing the mixing ratio, cross reaction with other pathogen positive serum of grouper is avoided, and the detection accuracy is ensured.

The invention provides a primer group, a kit and a detection method for detecting telosma mosaic virus (DelMV) in passion fruit leaves, and relates to a primer group for detecting the telosma mosaic virus (DelMV) in the passion fruit leaves and a kit for detecting the telosma mosaic virus (DelMV) in the passion fruit leaves. The sequences of the primer group designed based on a TelMV genome coat protein conserved region are shown as sequences 1-5 in a sequence table, a complete LAMP reaction system is established, and visual detection is realized by combining the primer group with a nucleic acid detection test strip. The method does not need complex instruments and professional operation, only needs a simple constant temperature device, and has the characteristics of rapidness, high specificity, high sensitivity and visual and easy result interpretation. By applying the method, virus-carrying seed trees can be removed in time, healthy seedlings can be guided to be cultivated, allocated and transported, technical support is provided for early diagnosis and monitoring of virus diseases, and the method has important significance on prevention and control of the virus diseases of the passion fruits in the field.

The invention provides a bivalent oral vaccine for fish, a preparation method and application, recombinant plasmids are electrically transferred into lactobacillus casei to prepare the bivalent oral vaccine for fish, the recombinant plasmids contain dominant antigen epitope sequences of I-type and II-type grass carp reoviruses, and the dominant antigen epitope sequences are shown as SEQ ID NO.1. The bivalent oral vaccine for fish is used for preparing the bivalent oral vaccine for fish. According to the bivalent oral vaccine disclosed by the invention, dominant antigen epitopes screened from I-type and II-type GCRV coat proteins are used as immunogens, so that the bivalent oral vaccine has special immunogenicity on the I-type and II-type GCRV; the screened dominant antigen epitope sequence has the advantages that the coded protein is small, and toxicity and harm to fish bodies are avoided. After entering a fish body, the bivalent oral vaccine for the fish can be expressed in the body and can start immune response, so that grass carp can be well protected from being infected by GCRV virus, and the survival rate is increased.

The invention discloses an RVVP4 / VP7 trimerized nano vaccine based on a ZIF-8 carrier as well as a preparation method and application of the RVVP4 / VP7 trimerized nano vaccine, and belongs to the technical field of biological medicines. According to the invention, the surface of the particle is modified with a CpG adjuvant through electrostatic interaction to form a pVAX1-coated ZIF-8-CpG compound, so that the protection of antigens and the targeted activation of dendritic cells (DCs) are realized; by constructing a DNA vaccine for coding RV coat proteins VP4 and VP7 and introducing an isoleucine zipper (IZ) and a T4 phage fibrin (Fd) trimerization structural domain, the stability and immunogenicity of antigen protein are remarkably enhanced, and a new strategy with remarkable competitive advantages is provided for development of RV vaccines. The invention not only overcomes the key bottlenecks that the traditional DNA vaccine is easy to degrade, poor in targeting and low in immunogenicity, but also provides a reliable technical platform with great transformation prospects for developing a new generation of RV vaccines through multiple strategies of'antigen rational design-nano targeting delivery-adjuvant synergistic immunization '.

The present application relates to the technical field of biology, in particular to a method for predicting influenza virus mutation, further relates to a method for demarcating the HA surface antigen region of influenza virus coat protein and the role in predicting influenza virus mutation. The method for predicting influenza virus mutation comprises: immunogenic region demarcation, prediction template selection, simulation mutation, screening and sequencing. The present application provides the feasibility of predicting the future mutation of influenza, and proposes a practical model to solve this problem. The present application can help to develop preventive therapeutic drugs in advance to better cope with the threat of influenza.

This invention provides adeno-associated virus (AAV) mutants and their applications. The capsid protein of the AAV mutant includes the amino acid sequences shown in SEQ ID NO:1-5, or amino acid sequences that have more than 98% identity with SEQ ID NO:1-5 and have the same or similar biological functions. This invention constructs an AAV library by splicing and recombinating the capsid protein encoding gene of wild-type AAV, and obtains AAV mutants by screening with neutralizing antibodies against the AAV capsid protein. These mutants exhibit strong infectivity against human liver cells and can evade the neutralizing effect of neutralizing antibodies, effectively addressing the technical obstacles in the treatment of hemophilia.

The present invention provides vector constructs comprising an open reading frame comprising a nucleic acid sequence encoding a modified pIX filamentous bacteriophage coat protein wherein the methionine (M) residue at position 1 of the pIX filamentous bacteriophage coat protein is replaced with a substitution amino acid residue. The invention also provides a bacteriophage particle or a bacteriophage display system containing the vector construct, and a bacteriophage display method and kit.

The present application belongs to the field of fish molecular immunology, and particularly relates to an oral vaccine containing a largemouth bass iridovirus ATPase and a preparation method thereof. B. subtilis The spores of the WB600 strain are antigen delivery carriers, the spore coat protein C (CotC) is used as an anchor protein, and the key protein of the largemouth bass iridovirus is fused with the ATPase gene and CotC the gene to construct a recombinant spore surface display system, so that the ATPase antigen is stably displayed on the spore surface, and an oral vaccine is prepared. The vaccine can tolerate the gastrointestinal environment after being orally administered to fish, and is accurately delivered to the gut-associated lymphoid tissue to stimulate the immunity of the fish, so that the prevention and control of the largemouth bass iridovirus is finally achieved.

The invention belongs to the technical field of biological vaccine preparation, and particularly relates to a preparation method of a targeted antigen displayed MS2 phage VLPs nanoparticle vaccine. The preparation method sequentially comprises the following steps: preparation and activation of a targeting ligand, fixed-point transformation and expression of MS2 bacteriophage capsid protein, preparation of targeting antigen-aptamer fusion protein, folding optimization and activity pre-verification of the antigen-aptamer fusion protein, self-assembly and antigen display of targeting MS2-VLPs, purification and characterization of targeting VLPs nanoparticles, and preparation of the vaccine preparation. On the basis, by introducing the targeting ligand aiming at the specific receptor on the surface of the antigen presenting cell, accurate recognition and combination of the VLPs vaccine on the target cell are realized, the problem that the antigen display lacks targeting is fundamentally solved, the antigen delivery efficiency is remarkably improved, the off-target effect is effectively reduced, and the antigen waste is avoided.

The application discloses a hybridoma cell strain secreting anti-Prunus necrotic ring-spot virus (PNRSV) monoclonal antibody and application of the monoclonal antibody. BALB / c mice are immunized by using a prokaryotic expression PNRSV coat protein (CP) as an antigen, and a hybridoma cell strain 9E1 capable of secreting anti-PNRSV monoclonal antibody is obtained by using a hybridoma technology, and the preservation number of the hybridoma cell strain is CGMCC No. 45329. The indirect ELISA titer of the monoclonal antibody secreted by the cell strain reaches 10 ‑7 , the antibody type and subclass are IgG2b and kappa light chain, the monoclonal antibody has specific immunoreaction with PNRSV, and does not have immunoreaction with Prunus pustule virus, cucumber green mottle mosaic virus, apple stem grooving virus and apple necrotic leaf virus infected leaf and healthy plant tissues. A plurality of detection methods are established by using the 9E1 monoclonal antibody, wherein the sensitivity of ACP-ELISA and dot-ELISA methods for detecting the diseased leaf reaches 1:10240 and 1:1280 times of dilution (w / v, g / mL) respectively. The cell strain is created, and the serological detection method is established, thereby providing detection technology and detection reagent support for PNRSV detection diagnosis, epidemiological investigation and scientific prevention and control.