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Pseudorabies virus SA215, pseudorabies virus polygene deletion bacterin and preparation method thereof

A pseudorabies virus, SA215 technology, applied in the field of animal virology and genetic engineering, can solve problems such as deletion, reduced virulence, and poor immunogenicity of vaccines

Active Publication Date: 2008-05-28
SICHUAN AGRI UNIV
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  • Abstract
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  • Claims
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Problems solved by technology

[0006] In order to overcome technical problems such as poor immunogenicity of existing vaccines, low safety, and difficulty in distinguishing wild virus-infected pigs from vaccine-immunized pigs, the present inventors used the Fujian A strain of pseudorabies virus as the original material, through homologous recombination Methods Several genes affecting virulence in Pseudorabies virus, TK, gE, gl, 11K, 28K, were artificially deleted, and a strain of SA215 virus with reduced virulence and no effect on the immunogenicity of Pseudorabies virus was obtained

Method used

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  • Pseudorabies virus SA215, pseudorabies virus polygene deletion bacterin and preparation method thereof
  • Pseudorabies virus SA215, pseudorabies virus polygene deletion bacterin and preparation method thereof
  • Pseudorabies virus SA215, pseudorabies virus polygene deletion bacterin and preparation method thereof

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Embodiment Construction

[0032] The construction of embodiment 1 pseudorabies virus gene deletion vaccine strain SA215

[0033] 1 Construction of pseudorabies virus gE / gI double gene deletion mutant strain:

[0034] 1.1 Construction of pPB7-1 plasmid

[0035] 1.1.1 Enzyme digestion of PRV Min A strain DNA

[0036] Virus PRV Min A strain comes from China Veterinary Drug Control Institute. PK15 cells were purchased from the Typical Microorganism Preservation Center of Wuhan University.

[0037] Cell culture and virus proliferation: PK15 cells that have overgrown a single layer were washed three times with calcium-magnesium-free water, digested with 0.25% trypsin for 2-10 minutes, added MEM nutrient solution containing 10% calf serum and kept at 37°C with 5% CO2 Cultivate in the incubator for 24-72 hours, after the cells grow into a dense monolayer, pour off the nutrient solution, add 0.2ml of PRV Min A strain virus solution, absorb at 37°C for 1 hour, add MEM containing 2% calf serum Maintenance sol...

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Abstract

The invention belongs to the technical field of animal virology and genetic engineering, relating to pseudorabies virus which lacks parts of virulence gene of min strain A of pseudorabies virus, and the process for preparing vaccine with the virus. The DNA recombinant technology is employed to artificially remove all or parts of albumen code area of a plurality of genes of TK, gE, gI, 11K, and 28k which are unnecessary genes and duplicate virus of viruses and influence the virulence in the pseudorabies virus, thereby lowering the virulence of the pseudorabies virus, achieving attenuated vaccine strains SA215 of which the attenuation genetic engineering is lack, preparing vaccine with SA215 virus, and immunizing animals to effectively prevent and control the occurrence and prevalence of the pseudorabies.

Description

Technical field: [0001] The invention belongs to the technical field of animal virology and genetic engineering, and relates to a pseudorabies virus lacking partial virulence genes of the Fujian A strain of pseudorabies virus and a method for preparing a vaccine with the virus. Background technique: [0002] Pseudorabies (PR) is a variety of domestic and wild animals caused by Pseudorabies Virus (PRV) of the Herpesviridae, α-herpesvirus subfamily, with fever, itching and encephalomyelitis as the main symptoms an acute infectious disease. Pigs are the primary infected host of this disease, and they are also the long-term storage and detoxification of the virus. The occurrence and prevalence of the disease have two main features: a large number of piglets within two weeks of age with neurological symptoms and death, and maternal reproductive disorders characterized by abortion, stillbirth and mummified fetuses or births in sows. Out of weak fetuses and stillbirths. At prese...

Claims

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Application Information

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IPC IPC(8): C12N7/00C12N15/33A61K39/245
Inventor 郭万柱
Owner SICHUAN AGRI UNIV
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