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A kind of pseudorabies virus gb-gc epitope tandem and application thereof

A technology of pseudorabies virus and porcine pseudorabies, applied in the direction of virus/bacteriophage, virus, virus peptide, etc., can solve the problem of failing to prevent the release of virus

Active Publication Date: 2022-05-17
INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Previous studies showed that the PRV-barthak61 vaccine failed to prevent viral shedding against virulent PRV strains in pigs (Wang et al., 2014; Tong et al., 2015)

Method used

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  • A kind of pseudorabies virus gb-gc epitope tandem and application thereof
  • A kind of pseudorabies virus gb-gc epitope tandem and application thereof
  • A kind of pseudorabies virus gb-gc epitope tandem and application thereof

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Experimental program
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Effect test

Embodiment 1

[0019] 1 Experimental materials and methods

[0020] 1.1 Reagent materials

[0021] FlyCut® EcoRI, FlyCut ® NCOI , EasyPure ® Quick Gel Extraction Kit, EasyPure ® Plasmid MiniPrep Kit (EM101), Agarose, GelStain, BL21(DE3) Chemically Competent Cell , ProteinRuler ® II (12-120 kDa), IPTG, His-tagged monoclonal antibody and horseradish peroxidase-labeled goat anti-rabbit or goat anti-mouse antibody were purchased from Beijing Quanshijin Co., Ltd. New Zealand white rabbits (2 kg) were purchased from Shanghai Jiaqian Biotechnology Co., Ltd.

[0022] 1.2 Amplification of gB and gC epitope genes

[0023] According to the PRV reference strain Becker sequence (JF79721 9.1) registered in GenBank, the primers for PCR amplification of the gB and gC antigenic epitope genes were designed, and the nucleic acid of the isolated new pseudorabies virus FJ-2012 strain was used as a template for PCR amplification. The augmented products were sent to Shenggong Biology (Shanghai) Engin...

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Abstract

The present invention provides a pseudorabies virus gB-gC epitope tandem and its application. By screening and cloning FJ-2012 strain gB and gC main antigen epitope genes, optimizing codons to form a tandem, artificially synthesizing its cDNA sequence, and Cloned into the prokaryotic expression vector pET-28a(+) multiple cloning site through NcoI and EcoRI restriction sites, transformed into BL21 (DE3) competent cells, the results of plasmid sequencing and restriction enzyme digestion showed the construction of gB-gC epitope tandem Gene recombination expression plasmid, gB-gC recombinant protein can be successfully expressed after IPTG induction for 2 hours, and the antiserum titer can reach 1:6400 as detected by Elisa. It indicated that the prokaryotic expression vector of PRV antigen tandem epitope gene was successfully constructed and PRV antiserum was obtained, which laid the foundation for the research of PRV epitope screening, PRV multi-epitope vaccine and diagnostic reagents.

Description

technical field [0001] The invention belongs to the technical field of animal husbandry and veterinary medicine, and in particular relates to a pseudorabies virus gB-gC epitope tandem and an application thereof. Background technique [0002] Porcine pseudorabies virus (PRV) is a porcine neuropathic herpesvirus (Ye et al., 2018). Although pigs are the natural host of PRV, it can also infect a variety of mammals, including ruminants, carnivores, and rodents. It is an important nervous system pathogen in livestock, and animals infected with PRV may die of central nervous system diseases (Tirabassi & Enquist, 2000 ). PRV infection poses a serious threat to the swine industry, and live-attenuated or inactivated vaccines are commonly used to control the disease (Freuling et al., 2017). Since 2011, pseudorabies caused by PRV gene mutation has reappeared in bartha-K61 vaccinated pig farms in China, and a new strain of PRV, FJ-2012, was isolated (Wu et al., 2017). Previous studies...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/62G01N33/569C12N15/70A61K39/245A61P31/22C12R1/19
CPCC07K14/005G01N33/56994C12N15/70A61K39/12A61P31/22C12N2710/16722G01N2333/032C07K2319/00A61K2039/552C12N2710/16734C12N2800/22
Inventor 吴学敏陈如敬陈秋勇王隆柏车勇良严山刘玉涛周伦江
Owner INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI