Bovine viral diarrhea virus (BVDV) transgenosis Astragalus mongholicus vaccine and production method

A technology of bovine viral diarrhea and virus genes, applied in biochemical equipment and methods, antiviral agents, genetic engineering, etc., can solve problems such as low protection, poor immunogenicity of nucleic acid vaccines, unsafe use of live virus vaccines, etc. To achieve the effect of no negative effects, fast gene homozygosity, and safe use

Inactive Publication Date: 2011-01-05
JILIN AGRICULTURAL UNIV
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  • Application Information

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Problems solved by technology

[0003] The purpose of the present invention is to provide a kind of bovine viral diarrhea virus transgenic astragalus vaccine, which is used for the prevention and treatment of animal bovine viral diarrhea, and overcomes the existing inactivated vaccines, nucleic acid vaccines with poor immunogenicity and low protection, and live virus vaccines are not suitable for use. security disadvantages

Method used

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  • Bovine viral diarrhea virus (BVDV) transgenosis Astragalus mongholicus vaccine and production method
  • Bovine viral diarrhea virus (BVDV) transgenosis Astragalus mongholicus vaccine and production method
  • Bovine viral diarrhea virus (BVDV) transgenosis Astragalus mongholicus vaccine and production method

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Embodiment Construction

[0076] 1. Bovine viral diarrhea virus gene E 0 Gene acquisition

[0077] Using the bovine viral diarrhea virus genomic RNA extracted by the conventional method of extracting RNA as a template, the reverse transcription-polymerase chain reaction was used to amplify E 0Gene. The primer sequences used for the amplification are as follows: upstream: 5′-CCGGATCCACCATGGAAAACATAACACAGTGG-3′; downstream: 5′-GCGAGCTCTTAAGCGTATGCTCCAAACCACGT-3′. After the product of reverse transcription-polymerase chain reaction was subjected to agarose gel electrophoresis, the target gene E 0 See figure 1 .

[0078] 2. Bovine viral diarrhea virus gene E 0 Construction of recombinant plant expression vector

[0079] Digest the gel-purified target gene E with DNA restriction endonuclease 0 and plant expression vector DNA, ligated with T4 DNA ligase at 16°C for 16 hours, took 5 μl of the ligated product to transform competent E. Identification of endonuclease double digestion and sequence analysi...

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Abstract

The invention relates to a bovine viral diarrhea virus (BVDV) transgenosis Astragalus mongholicus vaccine and a production method which are used for preventing and curing animal bovine viral diarrhea virus, and overcoming the defects of poor immunogenicity and low protection of the existing inactivated vaccines and nucleic acid vaccines as well as unsafe application of the existing live poison vaccine. In the invention, BVDV RNA is taken as a template, a reverse transcription-polymerase chain reaction method is used for obtaining an E0 gene thereof, then a gene E0 recombination plant expression vector is constructed, a pollen tube pathway method is used for obtaining the BVDV E0 gene transgenosis Astragalus mongholicus, and the Astragalus mongholicus with immunogen transformed by the BVDV E0 gene is screened by an immunoblotting and an enzyme linked immunosorbent assay to obtain a vaccine product. The invention has the following beneficial effects: the BVDV E0 gene is expressed in the Astragalus mongholicus, which generates the protective antigen and adjuvant substance capable of increasing the immunization, thereby improving the antiviral effect of the Astragalus mongholicus and the optimun immunity of the vaccine; and the invnetion has the advangtages of easy production, convenient use, strong stability and no side effects.

Description

technical field [0001] The invention belongs to the field of veterinary medicine technology or biotechnology, and in particular relates to a bovine viral diarrhea virus transgenic astragalus vaccine product and a preparation method. Background technique [0002] Bovine viral diarrhea virus is distributed worldwide, and there are many susceptible animals, such as deer, cattle, pigs, sheep, etc., and they can infect each other. The disease caused by this virus can occur year-round. Bovine viral diarrhea virus can cause various clinical symptoms such as viral diarrhea, mucosal disease, abortion, stillbirth, weak fetus, deformed fetus, persistent infection, immune tolerance, thrombocytopenia and hemorrhagic syndrome in animals, and can also cause Atypical swine fever has caused great harm to the deer industry, dairy industry and pig industry; especially the virus has been found in human vaccines, which is a potential threat to humans. Bovine viral diarrhea disease is a global ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12A61P31/14C12N15/83C12N15/40A61K36/481
Inventor 郜玉钢张连学臧埔李璠瑛王南李然杜锐王全凯刘佳佳于文影王亚兴
Owner JILIN AGRICULTURAL UNIV
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