Porcine pseudorabies mutant strain ge and gi gene deletion virus strain and its application

A technique for porcine pseudorabies and gene deletion, which can be used in antiviral agents, viruses/phages, biochemical equipment and methods, etc.

Active Publication Date: 2016-01-06
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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  • Porcine pseudorabies mutant strain ge and gi gene deletion virus strain and its application
  • Porcine pseudorabies mutant strain ge and gi gene deletion virus strain and its application
  • Porcine pseudorabies mutant strain ge and gi gene deletion virus strain and its application

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Experimental program
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Effect test

Embodiment 1

[0030] Embodiment 1: the acquisition of recombinant virus

[0031] 1.1 Viruses and Cells

[0032] 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.

[0033] Plasmids and Strains

[0034] 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); electroporation co...

Embodiment 2

[0056] 2.1 Rabbit body test

[0057] 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. The first and second groups were inoculated with F5PRV-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.

[0058] Preparation of Inactivated Vaccine and Pig Immune Protection Test

[0059] 2.2.1 Vaccine preparation

[0060] Take ZJ011G virus solution (10 7.0 TCID 50 / mL), inactivated with 0.1% formaldehyde at 37°C for 24 hou...

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Abstract

The present invention involves the field of pseudo-rabies virus technology. It specially involves a pseudo-pseudo-rabies disease variable and GI gene lack of virus strain PRV-ZJ011g. The preservation number is CGMCC?No.7957.Its application in the preparation of vaccines.Will 106.0?The TCID50 reorganization virus inocus the New Zealand big white rabbit, which did not cause itching and other clinical symptoms.Made into an oily vaccine, 4 weeks of GB after the piglets are immunized?ELISA antibody is positive, but GE antibody is negative, and immune protection efficiency is 100%. After the sow immunization, its offspring can obtain immune protection, which is 100%of PRV mutant poison and traditional toxic immune protection efficiency;Can be used for vaccine preparation.

Description

technical field [0001] The invention relates to the technical field of porcine pseudorabies virus, in particular to a porcine pseudorabies variant gE and gI gene deletion virus strain, and also relates to the application of the virus strain. Background technique [0002] Porcine pseudorabies virus ( Pseudorabies virus , PRV) is a member of the herpesviridae herpesvirus A subfamily, which can cause central nervous system disorders in newborn piglets, respiratory symptoms in growing pigs, and can cause abortion, stillbirth and weak piglets in pregnant sows. The United States and some European countries have declared the eradication and eradication of the disease. The gene-deficient attenuated vaccine is widely used in my country to effectively control and gradually purify it. [0003] Huazhong Agricultural University applied for a publication number CN1940063A in 2005, titled "A Pseudorabies E - / gI - Gene Deletion Strain, Inactivated Vaccine Containing the Strain, and App...

Claims

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Application Information

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Patent Type & Authority Patents(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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