Pseudo-rabies gE/gI-gene loss poison strain, killed vaccine containing it and use

A gene-deficient vaccine and pseudorabies virus technology, applied in the field of animal virology, can solve problems such as relapse, and achieve the effect of highly targeted prevention and high biological safety

Active Publication Date: 2007-04-04
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The applicant once developed a kind of pseudorabies gene vaccine (referring to Chinese Invention Patent Application Publication Specification, Patent Application No. 03156706.1) that has lost TK / gE / gI gene and has better immune effect and does not contain exogenous gene. There is a great contribution of the vaccine, but as a weakened live vaccine, there may still be a risk of returning to strength

Method used

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  • Pseudo-rabies gE/gI-gene loss poison strain, killed vaccine containing it and use
  • Pseudo-rabies gE/gI-gene loss poison strain, killed vaccine containing it and use
  • Pseudo-rabies gE/gI-gene loss poison strain, killed vaccine containing it and use

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0040] 1. Primer design

[0041] According to the published gene sequence of Prv Ea strain gI (gene database accession number is AF306511); gE (gene database accession number is AF171937) gene sequence, a pair of specific primers were designed and synthesized, abbreviated as P1 and P2, to amplify the double gene deletion of gI and gE The final DNA fragment size is 1400bp.

[0042] The primer sequences are as follows:

[0043] P1: 5'-TAGACGGGACGCTGCTGTTTCTGGAGG-3',

[0044] P2: 5'-CGGTCACGCCATAGTTGGGTCCATTCG-3'

[0045] The PCR amplification conditions are shown in Table 1:

[0046] Primer

PCR reaction conditions

PCR reaction condition system

E - / gI -

94°C 4min, 94°C 1min

51.8°C 1min, 72°C 1.5min

35cycles, 72℃10min

10×buffer (without Mg 2+ )5.0 μL,

Template 5.0μL

25mmol / L MgCl 2 8.0μL

2mmol / LdNTP 3.0μL

10mmol / L P1 2.0μL

10mm...

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Abstract

A recombinant Pseudorabies virus PrV gene engineering strain WKQ-001, inactivated vaccine containing the poisonous strain and its use are disclosed. It can be used to discriminate and diagnose artificial immunity pig or natural infectious pig. It's safe and doesn't contain exogenous gene.

Description

technical field [0001] The invention belongs to the technical field of animal virology and is related to genetic engineering. The present invention specifically relates to the construction of a recombinant pseudorabies virus (Pseudorabies virus, PrV) mutant strain (strain number: WKQ-001) and the use of the recombinant strain to prepare a pseudorabies gene deletion marker inactivated vaccine. The virus strain lacks the glycoprotein genes gI and gE of PrV, which leads to the reduction of PrV virulence, and thus can be used to prepare a pseudorabies safe vaccine. [0002] The invention also relates to the application of the recombinant pseudorabies virus strain and gene deletion inactivated vaccine. Background technique [0003] Pseudorabies virus (PrV) belongs to Herpesviridae a herpesvirus subfamily, has the characteristics of rapid growth on cell culture, strong neurotropism and latent infectivity (Roizmorn B, Desrosiers R, Flecbenstein...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01A61K39/12
Inventor 金梅林张松林宋云峰方六荣陈焕春
Owner HUAZHONG AGRI UNIV
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