20 results about "Antigen Gene" patented technology

This invention discloses a recombinant adenovirus vaccine targeting the Tp0326 antigen and its preparation method, comprising a recombinant adenovirus vector and the Tp0326 antigen expressed therein. The recombinant adenovirus vector uses a replication-defective human adenovirus type 5 (Ad5) as a backbone, inserting the full-length Tp0326 antigen gene of *Treponema pallidum* into the adenovirus shuttle vector pshuttle-IRES-rGFP-1, and obtaining it through homologous recombination with the adenovirus backbone plasmid pAdEasy-1. This invention, using a replication-defective Ad5 as a vector, provides a recombinant adenovirus vaccine with high safety and efficient expression of the full-length Tp0326 antigen. The expressed antigen has a natural conformation, retains the ECL4 immunodominant epitope, and is more likely to induce a specific immune response, resulting in stronger immunogenicity. The preparation method involves constructing a recombinant adenovirus vector through homologous recombination, packaging it in HEK293T cells, and purifying it by CsCl density gradient centrifugation to obtain a high-purity, high-titer recombinant adenovirus vaccine suitable for large-scale production. The recombinant adenovirus vaccine can be administered via intramuscular or intranasal injection.

The invention relates to a full-length sequence amplification primer, an amplification method and a three-generation sequencing method of an HLA-I antigen gene, and belongs to the field of gene detection. The HLA-I antigen gene is subjected to PCR (Polymerase Chain Reaction) amplification by using 8 pairs of characteristic amplification primers. And each gene can be subjected to high-yield specific amplification on a full-length sequence containing complete intron and exon regions only through one round of amplification and enrichment, and 7.8 Kb long-fragment genes can be captured once at most. The experimental deviation caused by simultaneous amplification of a single gene by using a plurality of pairs of segmented primers and the indirect error caused by gene splicing are effectively avoided. The method comprises the following steps: carrying out quality inspection and purification on amplicon PCR (Polymerase Chain Reaction) products, mixing samples, constructing an HLA-I type antigen gene library, carrying out accurate and complete sequencing on the full-length sequence of the HLA-I type antigen gene by adopting a three-generation PacBio Sequel II platform, and ensuring that the Hifi reads base accuracy can reach 99% or above. The reading of HLA-I type antigen gene full-length sequence variation information and the haplotype analysis at a high resolution level can be realized.

The invention relates to a recombinant rabies virus. The recombinant rabies virus is obtained by inserting an antigen gene of feline calicivirus and an antigen gene of feline herpes virus into a genome of a rabies virus attenuated strain. The triple inactivated vaccine for preventing and treating the feline calicivirus disease, the feline herpes virus disease and / or the rabies is prepared based on the recombinant rabies virus, and after the triple inactivated vaccine is used for immunizing cats, neutralizing antibodies aiming at the feline calicivirus disease, the feline herpes virus disease and the rabies virus can be effectively induced and generated; the simultaneous immunization on the three viruses is realized.

The application belongs to the technical field of biology, and discloses a preparation method of a Salmonella typhimurium flagellin and Senecavirus antigen fusion protein. The application selects Escherichia coli Rosetta (DE3) as a host, fuses a Salmonella typhimurium flagellin gene and a Senecavirus antigen gene, optimizes codons, and then clones the fusion protein into a pET-28a (+) vector, so that the obtained fusion protein can keep the adjuvant activity of the flagellin, improve the solubility and expression level of VP2, and the yield can reach 40 mg / L, thereby greatly reducing the expression and purification process cost. The fusion protein provided by the application can activate the TLR5 path and activate mouse macrophages, and the production process is simple and low in cost, so that the application has important significance for the research and production of Senecavirus subunit vaccines.

The application discloses a recombinant bacillus Calmette-Guerin (BCG) strain and application of the recombinant BCG strain in preparation of a tumor vaccine, wherein the recombinant BCG strain comprises an HPV16 E7 antigen gene with a nucleotide sequence as shown in SEQ ID NO:1; a cytolysin ClyA gene is further fused with the HPV16 E7 to be expressed, and a rBCG-ClyA+E7 recombinant strain is constructed; the recombinant BCG strain rBCG-E7 or rBCG-ClyA+E7 is applied in preparation of a vaccine for treating cervical cancer solid tumors; the anti-tumor effect is verified in a solid tumor model through a subcutaneous injection mode; the ClyA significantly improves the expression efficiency of an exogenous antigen in the BCG, and simultaneously enhances the presentation efficiency of an antigen presenting cell; the recombinant vaccine effectively activates an anti-tumor immune response, and does not cause obvious toxic side effects, thereby providing a safe and effective candidate strategy for immunotherapy of cervical cancer and other HPV related solid tumors.

According to one embodiment, a lipid particle for producing a CAR antigen-expressing cell derived from a CD209 positive cell is provided. The lipid particle includes a lipid membrane having a lumen, and a nucleic acid containing a CAR antigen gene being contained in the lumen.

The application belongs to the technical field of biotechnology, and provides a porcine deltacoronavirus recombinant DNA vaccine, a construction method and application thereof, the porcine deltacoronavirus recombinant DNA vaccine comprising a porcine deltacoronavirus NTD antigen gene or a porcine deltacoronavirus RBD antigen gene; wherein the nucleotide sequence of the porcine deltacoronavirus NTD antigen gene is shown as SEQ ID NO. 1, and the nucleotide sequence of the porcine deltacoronavirus RBD antigen gene is shown as SEQ ID NO. 2. The application firstly fuses the immunogenic gene fragments NTD and RBD of PDCoV with IgG1 Fc fragments respectively, constructs a recombinant DNA vaccine, and evaluates the safety, immunogenicity and protective efficacy of the recombinant DNA vaccine through experiments, and verifies that the PDCoV recombinant DNA vaccine has good effects in preventing porcine deltacoronavirus infection.

The invention establishes a method for simultaneously detecting vibrio parahaemolyticus, staphylococcus aureus, listeria monocytogenes, salmonella, shigella, bacillus cereus and vibrio alginolyticus on the basis of multiple PCR (Polymerase Chain Reaction) combined with a gene chip technology. The method comprises the following steps: selecting a heat-labile hemolysin gene of vibrio parahaemolyticus, a heat-resistant nuclease gene of staphylococcus aureus, a coded hemolysin O gene hlyA of listeria monocytogenes, an invasive protein A gene of salmonella enteritidis and an invasive plasmid antigen H gene of Shigella to carry out quintuple PCR (Polymerase Chain Reaction) amplification; bacillus cereus non-hemolytic toxin genes and vibrio alginolyticus similar Rpos sigma factor genes are selected for double PCR, two groups of multiplex PCR amplification products are hybridized with specific probes on the chip at the same time, and seven food-borne pathogenic bacteria can be detected at the same time. The method for detecting pathogenic bacteria by using the gene chip is high in specificity, high in sensitivity and strong in reliability.

The invention provides an engineered antigen, a gene, a cell and a reagent for detecting a rare blood group antibody, and application of the engineered antigen, the gene, the cell and the reagent. The engineered antigen for detecting the rare blood group antibody comprises a signal peptide, an antigen epitope sequence of rare blood group glycoprotein, a hinge region and a transmembrane region which are connected in sequence. The cell with the engineered antigen displayed on the surface can be used for rapid and accurate detection of the rare blood group antibody through a direct agglutination method, and the problems that reagent raw materials in an existing direct agglutination method are derived from human sources, are not easy to obtain, are difficult to store and the like can be solved. Compared with the prior art in which full-length antigen protein is expressed on cells, the method has the advantages that only the antigen epitopes of the rare blood groups are displayed on the surfaces of the cells, if antigens of different rare blood groups need to be replaced, only the antigen epitopes need to be replaced, the synthetic sequence is shorter, the method is simpler and more convenient, and the cost is lower.

The application discloses a liver fluke colloidal gold detection method based on specific antigen FHH, which adopts liver fluke specific antigen gene FHH, FHH protein, colloidal gold particles, S. aureus protein A labeled colloidal gold particles and a detection test strip to perform detection. The method has the advantages of simplicity and rapidness, visual result, low cost, high specificity and high sensitivity. The antigen gene FHH obtained through screening of a liver fluke adult cDNA expression library guarantees the specificity of the application. The application is suitable for clinical rapid diagnosis and large-scale epidemiological investigation.

The present invention aims to provide a method for immortalizing antibody-producing cells and the immortalized transformed cells obtained by this method. The present invention relates to a method for manufacturing immortalized cells, which involves introducing one or more genes selected from the group consisting of the SV40T antigen gene, the Bcl-2 family gene, and the Myc family gene and the Cyclin D family gene into non-immortalized antibody-producing cells. The present invention also relates to a method for manufacturing immortalized antibody-producing cells, which involves collecting antibody-producing cells from mammals, performing primary culture, and introducing one or more genes selected from the group consisting of the SV40T antigen gene, the Bcl-2 family gene, and the Myc family gene and the Cyclin D family gene into the obtained primary antibody-producing cells to create immortalized antibody-producing cells.

The invention provides a composition for preventing and / or treating an infection in an animal, comprising at least one antigen gene selected from the group consisting of SEQ ID NO: 1 (S273R), SEQ ID NO: 2 (E183L), SEQ ID NO: 3 (K78R), SEQ ID NO: 4 (KP177R), SEQ ID NO: 5 (A104R), SEQ ID NO: 6 (A137R), SEQ ID NO: 8 (B646R), SEQ ID NO: 10 (B602L), SEQ ID NO: 11 (CP2475L), SEQ ID NO: 12 (CP204L), SEQ ID NO: 13 (CP530R), SEQ ID NO: 14 (O61R), SEQ ID NO: 15 (D117L), SEQ ID NO: 16 (H108R The invention also provides methods of preventing and / or treating infections by administering the above compositions to an animal.

The invention belongs to the technical field of biology, and discloses a method for determining a multivalent antigen sequence of an anti-periodontitis red complex, which comprises the following steps: firstly, screening out key virulence factor antigens (FimA-II, RgpA, Msp and BspA) from three core pathogenic bacteria of the red complex: porphyromonas gingivalis, treponema denticola and fusisterone; then, independently cloning each antigen gene to a pUC57 cloning vector, constructing a single antigen recombinant plasmid, and avoiding intergene interference by designing a specific enzyme cutting site; and finally, through plasmid transformation, positive clone screening and double enzyme digestion verification, the plasmid quality is ensured. The obtained single antigen plasmid can be flexibly combined and used, is beneficial to subsequent multivalent vaccine development, and has the advantages of wide antigen coverage, high expression reliability, flexible construction strategy, easiness in industrialization and the like.

Vibrio parahaemolyticus serotype O10 is an important foodborne pathogen with a very high isolation rate in my country in recent years. This invention provides a specific oligonucleotide sequence for detecting Vibrio parahaemolyticus serotype O10 and its application. The oligonucleotides are DNA fragments selected from the IS1 and orf9 genes of the O antigen gene cluster of Vibrio parahaemolyticus serotype O10. This invention also establishes a PCR detection method for Vibrio parahaemolyticus serotype O10 using the above-mentioned oligonucleotides, establishing for the first time a technique for detecting this bacterium using molecular biology methods. This is of great significance for the identification and epidemiological monitoring of this important foodborne pathogen.

Provided are a fusion antigen of porcine Getah virus (GETV), a kit, a preparation method therefor and an application thereof. The fusion antigen of the GETV is primarily prepared by recombining a Gaussia luciferase (GLuc) gene with a codon- optimized GETV E2 antigen gene to construct an expression vector, and transfecting the expression vector containing GLuc-E2 into mammalian cell lines, resulting in the secretion of GLuc-E2 proteins into a cell supernatant for expression. Without the need for protein purification step, the cell supernatant may be directly collected for disease detection. The present disclosure demonstrates strong specificity and shows no cross- reactivity with African swine fever virus (ASFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), pseudorabies virus (PRV), or Japanese encephalitis virus (JEV).

This invention relates to a reverse genetics operating system for the segmented cloning and rescue of the Bartha K61 pseudorabies virus vaccine strain genome, its construction method, and its application. The system includes: recombinant plasmids BAC-cm-PRV-EA, BAC-cm-PRV-BC, and BAC-cm-PRV-FG containing the complete genome of the Bartha K61 pseudorabies virus vaccine strain, and host cells expressing the complete genome of the Bartha K61 pseudorabies virus vaccine strain. The reverse genetics operating system for the Bartha K61 pseudorabies virus vaccine strain of this invention successfully segments the Bartha K61 genome into six segments using ExoCET cloning and assembly technology, and then assembles them into three recombinant plasmids: BAC-cm-PRV-EA, BAC-cm-PRV-BC, and BAC-cm-PRV-FG. These three recombinant plasmids, after PacI linearization, can be co-transfected into African green monkey kidney cells to rescue the Bartha K61 virus, and the infectivity of the rescued Bartha K61 virus is comparable to that of the vaccine strain. Therefore, this reverse genetics operating system can provide an excellent vector for creating live vector vaccines related to the prevention and treatment of swine diseases, and can be used to develop Bartha K61 recombinant genetic engineering vector vaccines that express relevant swine disease antigen genes.

The application discloses a triplex forming oligonucleotide fluorescent probe and a detection method for detecting mycoplasma by targeting CARDS toxin genes, and belongs to the technical field of microbial molecular detection. The fluorescent probe is a triplex forming oligonucleotide (TFO), which is targeted to a specific and conservative core sequence of mycoplasma CARDS toxin genes, and a fluorescent reporter group is modified at the 5' end, and a fluorescent quenching group is modified at the 3' end. The TFO can form a stable triplex DNA complex with the target sequence through Hoogsteen base pairing, and trigger the release of a fluorescent signal. The detection method comprises sample pretreatment and DNA extraction, fluorescent probe incubation, fluorescent signal detection and result judgment in sequence, and does not need a PCR amplification step, and the whole detection process can be completed within 45 minutes. The probe takes a functional conservative region of the CARDS toxin gene as a target, effectively avoids detection deviation caused by variation of surface antigen genes, has a specificity of 100%, and has a detection lower limit of 1 copy / muL, and is suitable for mycoplasma detection in multiple scenes such as clinical treatment, biological pharmaceuticals and livestock breeding.

The present disclosure relates to methods for characterizing cancer testis antigen burden ("CTAB"), for predicting cancer survival outcomes using CTAB analysis, and for recommending and / or treating cancer using CTAB analysis. Particularly, aspects are directed to measuring expression of a panel of cancer testis antigen (CTA) gene markers in a tumor sample from a subject, determining a CTAB based on the measured expression of the CTA gene markers, classifying, by comparing to a reference CTAB score from the same cancer or tumor type, the CTAB score as a high-CTAB score or a low-CTAB score, and identifying the subject as responsive to a treatment wherein (i) the high-CTAB score is indicative of responsiveness to the treatment, or (ii) the low-CTAB score is indicative of responsiveness to the treatment. The cancer is preferably non-small cell lung cancer (NSCLC).

The invention relates to a three-generation library building and sequencing method for a full-length sequence of an HLA (Human Leukocyte Antigen)-III antigen gene, and belongs to the field of gene detection. According to the invention, PCR amplification is carried out on HLA-III type antigen genes by using five pairs of characteristic amplification primers. And each gene can be subjected to high-yield specific amplification on a full-length sequence containing complete intron and exon regions only through one round of amplification and enrichment, and 14Kb long-fragment genes can be captured once at most. The experimental deviation caused by simultaneous amplification of a single gene by using a plurality of pairs of segmented primers and the indirect error caused by gene splicing are effectively avoided. The HLA-III type antigen gene library is constructed by carrying out quality inspection and purification on amplicon PCR products and then carrying out sample mixing, the full-length sequence of the HLA-III type antigen gene is accurately and completely sequenced by adopting a three-generation PacBio Sequel II platform, and the Hifi reads base accuracy rate can reach 99% or above. The reading of HLA-III type antigen gene full-length sequence variation information and the haplotype analysis at a high resolution level can be realized.

The invention relates to a full-length sequence amplification primer, an amplification method and a three-generation sequencing method of an HLA-II antigen gene, and belongs to the field of gene detection. According to the invention, PCR amplification is carried out on HLA-II type antigen genes by using 10 pairs of characteristic amplification primers. Each gene can be subjected to high-yield specific amplification on a full-length sequence containing complete intron and exon regions only through one round of amplification and enrichment, and 17Kb long-fragment genes can be captured once at most. The experimental deviation caused by simultaneous amplification of a single gene by using a plurality of pairs of segmented primers and the indirect error caused by gene splicing are effectively avoided. The method comprises the following steps: carrying out quality inspection and purification on amplicon PCR (Polymerase Chain Reaction) products, carrying out sample mixing, constructing an HLA-II antigen gene library, carrying out accurate and complete sequencing on the HLA-II antigen gene full-length sequence by adopting a three-generation PacBio Sequel II platform, and ensuring that the Hifi reads base accuracy can reach 99% or above. The reading of HLA-II type antigen gene full-length sequence variation information and the haplotype analysis at a high resolution level can be realized.