30 results about "Bovine kidney" patented technology

The invention discloses a cell line used for separation culture and multiplication of an Orf virus (ORFV) as well as a preparation method and application of the cell line. The cell line disclosed by the invention is a newly born bovine testis supported cell line obtained by separating and purifying newly born bovine testis primary culture cells; research results show that the cell line is more sensitive to inoculation of contagious ovine ecthyma virus, virus titer multiplied by utilizing the cell line is about 1 virus titer higher than that of newly born bovine testis primary culture cells and 1.9 virus titer higher than that of a madin-darby bovine kidney (MDBK) cell line. ORFV is separated by utilizing the cell line, uniformity and stability of virus can be guaranteed, and a risk that the separated viruses carry other pathogens as the primary culture cells are utilized for multiple times can be prevented; besides, the passage cell line is utilized for preparing a vaccine, a production process of the vaccine can be simplified, a production cycle is shortened, production cost is reduced, and stable quality of the prepared vaccine is also guaranteed, so that the cell line has a certain practical significance on large-scale production of the vaccine.

The invention discloses the application of an andrographolide C15 substitution derivative in manufacturing an anti-hepatitis drug, which belongs to the technical field of pharmaceutical chemistry. According to the invention, MDBK (mardin-darby bovine kidney cells) and BVDV (bovine viral diarrhea virus) are utilized to study the inhibitory action of the compound on cytopathic effect caused by the virus through an external screening model. Through the screening of a large number of andrographolide derivatives, the compound with a structure of general formula 1 is found to significantly inhibit MDBK cytopathic effect caused by the BVDV; the andrographolide C15 substitution derivative has high efficiency and low toxicity, thereby being used for manufacturing medicaments for treating and preventing hepatitis and having good prospect in development and application. The general formula 1 is shown in the description.

The invention discloses a recombinant porcine alpha interferon, prepared by the following methods: A1, synthesizing an artificially modified porcine alpha interferon; A2, constructing a recombinant eukaryotic expression vector pGAPZ alpha-IFN alpha; and A3, highly expressing IFN alpha by an eukaryotic cell yeast expression system. The invention is characterized by taking a supernatant to purifying the porcine alpha interferon which is cultured by a lot of engineering bacteria and expressed with a chromatography column, then collecting a target protein eluate, filtering through a 0.22 mum microfiltration membrane, then determining the activity by cytopathic inhibition, and calculating the potency unit according to 50% pathology; wherein the cell used for determination is Madin-Darby bovine kidney (MDBK), and vesicular stomatitis virus (VSV) is used for attacking the virus. According to the invention, the recombinant porcine alpha interferon obtained by purification is applied in controlling PCMV, and experiments prove that the recombinant porcine alpha interferon has obvious protection effect on PCMV infected cells. The route of administration of the porcine alpha interferon is injection or mucous membrane administration, and the dosage form comprises injection or nasal drops.

The invention discloses an infectious bovine rhinotracheitis virus strain and a preparation method and application thereof. A wild strain of infectious bovine rhinotracheitis virus is found. The preparation method comprises the following steps of: firstly, identifying a sample by a PCR (Polymerase Chain Reaction) and a fluorescence PCR; then adding bovine serum which is detected to be free of IBRV (Infectious Bovine Rhinotracheitis Virus) to MDBK (Madindarby Bovine Kidney) cells by means of a culture medium, cultivating for a certain period of time so as to grow single-layer cells; after inoculating the single-layer cells with the processed sample, observing the cytopathic effect; after taking a pathological cell sample and identifying the pathological cell to be positive by the PCR, measuring TCID50 (Tissue Culture Infective Dose) as 106.2, and carrying out the virus neutralization test, wherein the TCID is 27; then inoculating the sample into the single layer cells; and when 70 percent to 80 percent of cells undergo the cytopathic effect and storing the cells into a refrigerator at a temperature of -40 DEG C. The infectious bovine rhinotracheitis virus strain and the preparation method disclosed by the invention can be applied to preparation of an engineered vaccine for preventing the infectious bovine rhinotracheitis.

The invention belongs to the technical field of veterinary biological products, and particularly relates to swine-borne BVDV-2 strain infectious cDNA (complementary deoxyribonucleic acid) clone and application thereof. The swine-borne BVDV-2 strain infectious cDNA clone contains swine-borne BVDV-2 seed virus full-length cDNA, T7 RNA (ribonucleic acid) polymerase promoters are inserted in 5' terminals of the swine-borne BVDV-2 seed virus full-length cDNA, and SbfI restriction enzymes are inserted in 3' terminals of the swine-borne BVDV-2 seed virus full-length cDNA. The invention further discloses a plasmid pASH28 with the swine-borne BVDV-2 strain infectious cDNA. The plasmid is linearized and then is subjected to in-vitro transcription to obtain RNA, MDBK (Madin-Darby bovine kidney) cells are transfected by the plasmid, and bovine viral diarrhea viruses can be successfully rescued. The swine-borne BVDV-2 strain infectious cDNA clone, the application and the plasmid have the advantage that the swine-born BVDV-2 strain infectious cDNA clone can be applied to research on functional difference between different animal-borne BVDV-2 proteins and also can be used for genetically modifying and preparing high-titer attenuation BVDV vaccine.

The invention provides a caprine herpesvirus I vaccine strain and an application thereof, and relates to the field of biological veterinary products. The caprine herpesvirus I vaccine strain is a caprine herpesvirus I JSHA1405 strain, which is preserved with preservation number of CCTCC NO:V201745. Virus liquid is obtained through multiplication of the virus (caprine herpesvirus I) by virtue of mardin-darby bovine kidney cells. The invention also provides an application fot eh vaccine strain in preparing caprine herpesvirus I vaccines. The caprine herpesvirus I JSHA1405 strain provided by theinvention is relatively strong in pathogenicity and is good in immunogenicity. An oil adjuvant inactivated vaccine, which is prepared by the caprine herpesvirus I JSHA1405 strain, is safe and reliable; goats, which are immunized, can generate an antibody at a relatively high level, and the duration of the antibody can reach 6 months; in addition, an excellent protective effect can be taken on homologous virus seed toxicity attack; morbidity in the immunized goats is obviously reduced; and the prevalence of the caprine herpesvirus I can be effectively prevented.

The invention provides an anti-IBRV single-chain antibody which is protein consisting of a heavy chain variable region shown as SEQ ID NO.1, a light chain variable region shown as SEQ ID NO.2 and a connecting peptide for connecting the two regions and having the characteristic of resisting an infectious bovine rhinotracheitis peplos gD protein antibody. The invention also provides a preparation method of the single-chain antibody. The single-chain antibody can be specifically combined with IBRV and can be also specifically combined with IBV gD protein in an escherichia coli expression product. An indirect immunofluorescence assay shows that the single-chain antibody can identify the IBRV infected in the bovine kidney cell MDBK, and the single-chain antibody has the good application value when applied to infectious bovine rhinotracheitis detection and development of treatment preparations.

The invention discloses application of fructus chebulae in preparing a medicine for inhibiting and killing BVDV (bovine viral diarrhea virus). The application of the fructus chebulae in preparing themedicine for inhibiting and killing the BVDV has the beneficial effects that the Tibetan medicine is used as a study object, and the screened fructus chebulae water extract has certain inhibition on the BVDV in a MDBK(Madin-darby-Bovine-Kidney) model; the direct killing function on viruses is better than both the adsorption blocking function and the duplicating blocking function, and a foundationis laid for the preventing and controlling of BVD diseases and the study and development of medicines; the novel pharmacologic action of the fructus chebulae is found, the fructus chebulae can be usedas a candidate medicine for preventing and controlling the BVDV and/or clearing the infection of BVDV, and the fructus chebulae can be singly used or jointly used.

The invention discloses a method of screening MDBK (Madin-Darby Bovine Kidney) cell lines sensitive to bovine viral diarrhea/mucosal viruses. The method includes: performing mono-cloning on MDBK cells; inoculating bovine viral diarrhea/mucosal viruses of two biotypes (cytopathic and noncytopathic); adopting indirect immunofluorescence technology to measure viral titer, and determining the cell lines with the viral titer higher than 106TCID50/ml as the MDBK cell lines sensitive to the bovine viral diarrhea/mucosal viruses. The MDBK cell lines screened by the method are sensitive to the bovine viral diarrhea/mucosal viruses of the two biotypes, and the method can be directly applied in research and production of bovine viral diarrhea/mucosal disease vaccines.

The invention relates to the technical field of health-care products, and discloses oral liquid sea cucumber and cordyceps sinensis peptide, which is prepared from the following raw materials in percentage by weight: 25 to 35 percent of sea cucumber, 7 to 13 percent of fructus lycii, 3 to 7 percent of bovine kidney, 3 to 7 percent of radix ginseng, 3 to 7 percent of radix ophiopogonis, 0.5 to 2 percent of pericarpium citri reticulatae, 0.1 to 0.5 percent of xylooligosaccharide, 0.15 to 0.05 percent of sodium carboxymethyl cellulose, 0.15 to 0.05 percent of potassium sorbate, 0.1 to 0.3 percent of cordyceps sinensis and the balance of water. The traditional Chinese medicine formula has the advantages that the traditional Chinese medicine formula can tonify qi and yin, the traditional Chinese medicine liquid is used for nourishing, the activity of sea cucumber peptide can be well preserved, the sea cucumber peptide can be absorbed within one minute on an empty stomach, the sea cucumber peptide can be directly absorbed into blood and tonify the kidney without digestion, high-quality protein which is higher in quality and easier to absorb is provided for a human body, and the traditional Chinese medicine composition is more reasonably and effectively utilized; according to the formula, the sea cucumber peptide is processed and produced by adopting a biological enzymolysis technology, and other medicinal materials are processed and produced by adopting a multi-stage separation and high-pressure extraction technology.

The invention discloses an I-type interferon receptor (IFNAR) gene knockout bovine kidney cell line, which is obtained by carrying out gene editing on bovine kidney cells by utilizing a CRISPR/Cas9 gene editing technology and carrying out monoclonal selection. Phenotype verification shows that after the IFNAR1 knockout bovine kidney cell line does not express IFNAR1 and is infected with a bovine viral diarrhea virus (BVDV), an infectious bovine rhinotracheitis virus (IBRV) and a 3-type bovine parainfluenza virus (BPIV-3), the virus titer can be remarkably improved, so that the IFNAR1 knockout bovine kidney cell line can be used for separating and identifying bovine-derived virus clinical strains and improving the virus proliferation efficiency.

The present invention pertains to a Madin-Darby bovine kidney (MDBK) cell, wherein the interferon regulatory factors (IRF) IRF3 and/or IRF7 encoding genes are functionally inactivated. The invention also pertains to a cell culture comprising the MDBK cell, use of the MDBK cell culture, a method for the production of a virus using the cell and a vaccine prepared by using the cell.

The invention discloses a porcine BVDV (Bovine Viral Diarrhea Virus) recombinant virus for expressing porcine epidemic diarrhea virus (PEDV) S genes and the application thereof. A neutralization antigenic epitope sequence of the PEDV S genes is inserted into a terminal N of a C nucleocapsid protein gene of porcine BVDV. A recombinant infectious cloning plasmid is obtained. The plasmid is linearized, subjected to in vitro transcription to obtain RNA and transfected with MDBK (Madin-Darby Bovine Kidney) cells, so that the porcine BVDV recombinant virus for expressing the PEDV S proteins can be successfully rescued. The porcine BVDV recombinant virus is capable of expressing a PEDV S foreign protein, and is simultaneously used for controlling the PEDV and BVDV. The porcine BVDV infectious clone provides suitable insertion sites for other exogenous genes, can serve as a recombinant viral vector, and provides an effective technology platform for researches such as subsequent in-vitro modification of RNA, insertion of exogenous genes, preparation of recombinant viruses and the like.

A method for promoting replication of infectious bovine rhinotracheitis viruses using cold atmospheric plasma, including: irradiating a medium for Madin-Darby bovine kidney cells with a cold atmospheric plasma generator; adding the irradiated medium to the Madin-Darby bovine kidney cells; and adding infectious bovine rhinotracheitis viruses for incubation. The time required for treatment is 2 min allowing for a simple and rapid operation. The plasma is used for the indirect treatment of cells with a uniform process and a controllable intensity. The replication of the infectious bovine rhinotracheitis viruses in the Madin-Darby bovine kidney cells is significantly promoted by co-incubation in the treated DMEM for 1 hour, so that the high levels of infectious bovine rhinotracheitis viruses obtained can be used for vaccine production after inactivation, improving the vaccine production efficiency.

The invention discloses a shRNA (short hairpin Ribonucleic Acid) expression vector for interfering BVDV (Bovine Viral Diarrhea Virus) copying and a constructed recombinant cell. Different shRNA fragments are designed specific to different regions of a BVDV. SiRNA (small interfering Ribonucleic Acid) is formed by transcribing shRNA in a host cell and shearing, and acts on different regions of a virus RNA to interfere the copying and expression of viruses. BVDV cell surface receptors exists in bovine kidney cells, testicle cells and the like, so that BVDV is most sensitive to fetal bovine kidney cells and testicle cells, a complete transgenic cloned cattle individual is formed by performing induced rearrangement on a fetal bovine fibroblast serving as a somatic cell nuclear transplanting donor cell, and BVDV sensitive organs such as kidney and the like have the capabilities of suppressing virus expression.