TK, ge and gT gene deletion strain of porcine pseudorabies virus variant and application of TK, ge and gT gene deletion strain

A porcine pseudorabies virus, gene deletion technology, applied in the direction of viruses, viral peptides, antiviral agents, etc., can solve problems such as abortion and stillbirth of pregnant sows

Inactive Publication Date: 2017-06-13
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

However, since 2011, many pig farms immunized with live PRV vaccines in my country have had outbreaks of the disease again, mainly manifested as neurolog

Method used

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  • TK, ge and gT gene deletion strain of porcine pseudorabies virus variant and application of TK, ge and gT gene deletion strain
  • TK, ge and gT gene deletion strain of porcine pseudorabies virus variant and application of TK, ge and gT gene deletion strain
  • TK, ge and gT gene deletion strain of porcine pseudorabies virus variant and application of TK, ge and gT gene deletion strain

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Embodiment Construction

[0044] The present invention is further illustrated by the following examples.

[0045] 1 Materials and methods

[0046] 1.1 Main materials

[0047] 1.1.1 Viruses and Cells

[0048] PRV ZJ01 strain virus, which was isolated by our laboratory in February 2012 from sick piglets in a large-scale pig farm in Zhejiang, is a highly pathogenic strain and is adapted to the F8 generation of BHK-21 cells; PRV ZJ01ΔgE / gI strain was used in this experiment The gI and gE double-gene deletion virus obtained by using the PRV ZJ01 strain as a parent was adapted to the F10 generation of BHK-21 cells. BHK-21 cells were preserved by our laboratory, the cell growth medium was DMEM (Gibco) containing 10% fetal bovine serum (Gibco), and the maintenance medium was DMEM containing 2% fetal bovine serum.

[0049] 1.1.2 Plasmids and strains

[0050] pHA2 plasmid (including BAC vector sequence, gfp expression cassette expressed in eukaryotic cells, chloramphenicol resistance gene expressed in Escher...

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Abstract

The invention discloses a TK, ge and gT gene deletion strain of a porcine pseudorabies virus (PRV) variant ZJ01 and application of the TK/ ge/gT gene deletion strain. The strain is the TK/ ge/gT gene deletion strain ZJ01delta TK/gE/gI (ZJ01R in short) of the PRV ZJ01 strain. According to the in-vitro passage test results, gE/gI gene deletion strains ZJ01delta gE/gI of a recombinant virus ZJ01R and the PRV ZJ01 strain are consistent with cytopathic effect generated by a parental virus ZJ01, and virus titer is similar with the ZJ01 strain; a target gene deletion region of the recombinant virus ZJ01R continuously and stably exists without resilience mutation; according to the pig immunity and attacking protection test, after the recombinant virus ZJ01R is immune, a pig is free of clinical symptoms and pathological changes; after being immunized by 21 days, the pig can generate a PRV neutralizing antibody with relatively high level, and can resist challenging infection of a fatal PRV variant strain ZJ01.

Description

[0001] Technical field: [0002] The invention relates to a porcine pseudorabies virus variant strain used for preparing a vaccine, in particular to a porcine pseudorabies virus variant strain TK, gE and gI gene deletion strain. [0003] Background technique: [0004] Porcine pseudorabies is an important infectious disease of pigs caused by pseudorabies virus (PRV). The main clinical symptoms include: central nervous system disorders in newborn piglets, respiratory symptoms in weaned piglets and finishing pigs, and reproductive disorders in pregnant sows. , has caused huge economic losses to the world's pig farming industry [1-3] . At present, the prevention and control of the disease mainly relies on vaccine immunization. The United States and some European countries have announced that the epidemic of the disease in domestic pigs has been eradicated through gene deletion vaccine immunization and purification technology [4] . In the past 30 years, my country has widely used...

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Application Information

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IPC IPC(8): C12N7/00C12N15/85A61K39/245A61P31/22C12R1/93
CPCC07K14/005C12N7/00C12N2710/16721C12N2710/16722C12N2710/16734C12N2710/16751
Inventor 姜平董静顾真庆白娟王先炜李玉峰
Owner NANJING AGRICULTURAL UNIVERSITY
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