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Recombinant porcine pseudorabies virus strain and preparation method thereof

A technology of porcine pseudorabies virus and recombinant virus, which is applied in the field of virology and can solve the problems of few reports on live virus vector vaccines

Inactive Publication Date: 2014-07-30
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There are currently PCV2 inactivated vaccines, PCV2 DNA vaccines, subunit vaccines and chimeric virus vaccines in the research of PCV2 vaccines, but there are relatively few reports on live virus vector vaccines

Method used

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  • Recombinant porcine pseudorabies virus strain and preparation method thereof
  • Recombinant porcine pseudorabies virus strain and preparation method thereof
  • Recombinant porcine pseudorabies virus strain and preparation method thereof

Examples

Experimental program
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Embodiment Construction

[0037] Experiment 1 Construction of recombinant porcine pseudorabies virus transfer plasmid expressing porcine circovirus type II ORF2 gene and porcine IL-18 gene

[0038] 1 Materials and methods, 1.1 Materials, 1.11 Plasmids, viruses, cells and strains

[0039] The recombinant pseudorabies virus transfer plasmid PGO containing the PCV2 ORF2 gene and the recombinant plasmid pGEMT-IL-18 containing the porcine IL-18 gene were purchased from the Key Laboratory of Animal Food Safety in Henan Province. Porcine pseudorabies PRV TK - / gG - / gE - The vaccine strain was purchased from Wuhan Keqian Biological Products Company. Pig testis (ST) cells were purchased from China Veterinary Drug Administration (frozen storage); Escherichia coli (E.coli) JM109 was purchased from GibcoBRL.

[0040] 1.1.3 Main reagents DNA Marker λ-EcoT14I, DNA Marker Tans15K, DNA Marker DL2000, Premix Ex Taq DNA polymerase and restriction endonucleases BglII, BamHI, HindIII, etc. were purchased from Treasur...

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Abstract

The invention discloses a recombinant porcine pseudorabies virus strain (belonging to Herpesviridae) and a preparation method thereof. The recombinant porcine pseudorabies virus strain has the accession number of CCTCC NO: V201345. Based on porcine pseudorabies virus vectors, green fluorescent protein labels are used so that green fluorescent protein expression gG-less universal transfer vector PG is constructed, and the recombinant porcine pseudorabies virus strain contains a pseudorabies virus self-late gene gG promoter, a CMV promoter and SV40Poly(A), has upstream flanks of 0.8kb and downstream flanks of 1.7kb and completely satisfies homologous recombination demands. Enhanced green fluorescent protein (EGFP) can be used as a gene expression indicator and comprises porcine IL-18 gene inserted into multiple cloning sites of the EGFP so that connection construction of different expression cassettes by a common vector is realized and two expression cassettes are individually expressed. The exogenous gene expression quantity is closely related to the connection direction of the two expression cassettes.

Description

technical field [0001] The invention relates to the technical field of virology, in particular to a recombinant porcine pseudorabies virus expressing porcine circovirus type 2 ORF2 gene and porcine IL-18 gene and a preparation method thereof. Background technique [0002] Porcine circovirus (PCV) is one of the smallest animal viruses discovered so far. According to the pathogenicity, antigenicity and nucleotide sequence of PCV, PCV can be divided into two genotypes, PCV1 and PCV2. PCV1 widely exists in pigs and pig-derived cell lines. It is non-pathogenic and cannot cause cytopathic changes. PCV2 is pathogenic and mainly causes postweaning multisystemic wasting syndrome (PMWS) in weaned piglets. Various diseases caused by PCV2 infection in pigs have spread all over the world, bringing huge losses to the world's pig industry. Porcine circovirus type 2 has two main open reading frames ORF1 and ORF2, wherein the ORF2 gene is the main structural protein gene of PCV2, encoding ...

Claims

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Application Information

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IPC IPC(8): C12N7/01C12N15/63C12R1/93
Inventor 陈红英郑兰兰崔保安郭晓庆魏战勇朱前磊
Owner HENAN AGRICULTURAL UNIVERSITY
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