30 results about "Bovine kidney" patented technology

The invention discloses a health wine and a preparation method thereof, wherein the health wine comprises the following raw materials: bovine kidney, sheep kidney, penis et testis serpentis, cinnamon, polygonatum, raspberry, licorice, clove, cordyceps sinensis, yam, oyster, papaya, Chinese dates, wolfberry fruit, arillus longan, honey, crystal sugar and 30-degree Daqu spirit. The preparation method is as follows: cleaning the bovine kidney, sheep kidney, penis et testis serpentis, cinnamon, polygonatum, raspberry, licorice, clove, cordyceps sinensis, yam, oyster, papaya, Chinese dates, wolfberry fruit and arillus longan, adding water for leaching, filtering to separate filtrate liquor from filter residue, then repeatedly leaching twice, mixing filtrates obtained at the three times, heating and concentrating to obtain extract, mixing the extract, refined honey, refined crystal sugar and 30-degree Daqu spirit and evenly stirring, sealing and standing for one month, then taking out, filtering to remove impurities, and finally filling to obtain the finished product. The health wine has the efficacies of invigorating premodial energy, tonifying kidney yang, enriching essence and blood,supplementing blood, nourishing yin, and nourishing blood to tranquillize mind.

The present invention provides a plasmid containing a beef cattle DKK3 gene 3'UTR sequence and a dual luciferase reporter gene and application thereof, the plasmid contains a target for binding bta-miR-25 and bovine DKK3, and its construction includes designing a DKK3 gene The 3'UTR region amplification primers were added with enzyme cleavage sites Pme I and Xho I, and the bovine kidney cell (MDBK) cDNA was used as a template for amplification, and the PCR product and pmirGLO Vector were respectively used with restriction enzymes Pme I and Xho 1 carry out double digestion, link the carrier pmirGLO Vector and DKK3 gene 3'UTR fragment purified after the digestion, transform, cultivate, PCR detection, sequencing, the bacterial liquid that the sequencing is correct is re-expanded and cultivated, and steps such as plasmid extraction, the present invention It provides a simple and easy tool for screening the miRNAs that regulate the expression of DKK3 and evaluating the regulation effect of miRNAs on the expression of DKK3, and has the advantages of high sensitivity, fast speed and low cost; the present invention also brings deeper insights into the regulation of DKK3 expression. Cognition can be applied to the study of the molecular regulation mechanism of the genetic improvement of livestock meat quality, and has a good application prospect.

The invention discloses the application of bovine kidney cells capable of suspension culture in virus culture and vaccine preparation. The bovine kidney cell MDBK suspension culture method of the present invention saves a large number of manual operations such as cell digestion, liquid replacement, frequent sorting, etc., the culture method is simple and fast, and the reactor culture is used to realize process automation, which not only improves the cell yield per unit volume , It also solves the technical problem of differences between batches of traditional adherent cultured cells.

The invention relates to a construction method of recombinant coatpox virus used for expressing foot and mouth disease virus empty capsid. The method comprises the following steps of: constructing transfer plasmids on the basis of forward and reverse homologous arms; inserting a foot and mouth disease virus protein precursor gene P1-2A, a vaccinia virus back-to-back promoter P7.5-P11, a hypoxanthine-guanine phosphoribosyltransferase gene, a foot and mouth disease virus internal ribosome entry site sequence and a foot and mouth disease virus 3C protease 9-site mutant gene to the transfer plasmids in sequence; co-transfecting an MDBK (Madin-darby Bovine Kidney) cell through the transfer plasmids and the coatpox virus; and then inserting an exogenous gene between the forward and reverse homologous arms in a homologous recombination way. By adopting the construction method, the screened recombinant coatpox virus can exist stably and can be used for expressing the foot and mouth disease virus empty capsid.

The invention discloses bovine kidney cells capable of suspension culture and a suspension culture method and application thereof. The bovine kidney cell MDBK suspension culture method of the present invention saves a large number of manual operations such as cell digestion, liquid replacement, frequent sorting, etc., the culture method is simple and fast, and the reactor culture is used to realize process automation, which not only improves the cell yield per unit volume , It also solves the technical problem of differences between batches of traditional adherent cultured cells. The bovine kidney cell MDBK and the suspension culture cell thereof of the present invention can be used in fields such as the cultivation of various sensitive viruses, the production of antibody proteins, the production of vaccines, the inspection of exogenous viruses in products, and the like.

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.