gE- and gI-deleted porcine pseudorabies virus variant strain and use thereof

A technique for porcine pseudorabies and gene deletion, applied in the directions of antiviral agents, viruses/phages, biochemical equipment and methods, etc.

Active Publication Date: 2014-04-30
JIANGSU NANNONG HI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the problems such as the porcine pseudorabies virus vaccine in the above prior art can not resist the infection of highly virulent antigen variant epidemic strains, the invention provides a kind of can be used for preparing Porcine pseudorabies gE and gI gene deletion virus strain with high immune efficiency vaccine

Method used

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  • gE- and gI-deleted porcine pseudorabies virus variant strain and use thereof
  • gE- and gI-deleted porcine pseudorabies virus variant strain and use thereof
  • gE- and gI-deleted porcine pseudorabies virus variant strain and use thereof

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Experimental program
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Effect test

Embodiment 1

[0034] Embodiment 1: the acquisition of recombinant virus

[0035] 1.1 Viruses and cells

[0036] PRV-ZJ01 strain virus (identified as a highly virulent antigenic variant strain) was isolated by our laboratory in 2011 from a sick piglet in a certain pig herd in my country, and it is an adaptive virus for the F5 generation of BHK-21 cells; The cell growth medium was DMEM (Gibco) containing 10% fetal bovine serum (Gibco), and the maintenance medium was DMEM containing 2% fetal bovine serum.

[0037] Plasmids and Strains

[0038] pHA2 plasmid (contains BAC vector sequence, expressed in eukaryotic cells gfp The expression cassette, chloramphenicol resistance gene expressed in Escherichia coli and Escherichia coli F plasmid) were donated by Dr. Wang Jichun, National Veterinary Biological Products Engineering Technology Research Center; pUC19 vector was purchased from Invitrogen; Escherichia coli TOPO10 competent cells (provided by our laboratory preservation); electroporat...

Embodiment 2

[0060] 2.1 Rabbit body test

[0061] Nine New Zealand white rabbits (Experimental Animal Center, Jiangsu Academy of Agricultural Sciences) were randomly divided into 3 groups, 3 rabbits in each group, and kept in isolation. Groups 1 and 2 were inoculated with F5 PRV-ZJ01 and ZJ011G respectively, subcutaneously inoculated on the back, 10 6.0 TCID 50 / mL, the third group was used as a blank control, and observed whether clinical symptoms such as itching occurred every 8 hours. 3-5 days after PRV-ZJ01 inoculation, all 3 rabbits had obvious itching symptoms and died, and PRV was present in the brain tissue test, while the ZJ011G and blank control rabbits had no obvious symptoms, which proved that the virulence of ZJ011G was significantly reduced.

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Abstract

The invention relates to the technical field of porcine pseudorabies viruses and especially relates to a gE- and gI-deleted porcine pseudorabies virus variant strain PRV-ZJ011G and a use thereof. The gE- and gI-deleted porcine pseudorabies virus variant strain PRV-ZJ011G has the accession number of CGMCC No.7957. The invention discloses the use of the gE- and gI-deleted porcine pseudorabies virus variant strain PRV-ZJ011G in vaccine preparation. After the New Zealand big white rabbit is inoculated with the 106.0TCID50 recombinant viruses, clinical symptoms such as pruritus are not caused. An oil-in-water inactivated vaccine prepared from the gE- and gI-deleted porcine pseudorabies virus variant strain PRV-ZJ011G is injected into a piglet and after four weeks, the BELISA antibody is produced but the gE antibody does not exist, and the immunization protection efficiency is 100%. After immunization on sows, the piglets produced by the sows get immunization protection and the efficiency of PRV variant virus and traditional virus immunization protection is 100%. It is proved that the ZJ011G recombinant virus has good immunogenicity and can be used for vaccine preparation.

Description

Technical field [0001] The present invention involves the field of pseudo -rabies virus technology, especially involving a pseudo -puppet rabies degenerative strain and GI gene lack of virus strains, and the application of the virus strain. Background technique [0002] Puppet rabies virus ( Pseudorabies virus , PRV) is a member of the Asian Department of herpes virus -type herpes virus, which can cause disorders of the new nervous system of piglets, growing pig respiratory symptoms, and can cause pregnancy sow abortion, dead tires and weak babies.The United States and some European countries have announced the elimination and eradication of the disease.my country is widely used in China to effectively control and gradually purify the lack of weak toxic vaccines. [0003] Huazhong Agricultural University applied for a public number CN1940063A in 2005, named "a pseudo -rabies disease E - / gi - Gene lack of strains, analogy and applications containing the strain ", and the pseudo-...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01A61K39/245A61P31/22
Inventor 姜平顾真庆王继春白娟
Owner JIANGSU NANNONG HI TECH
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