Recombinant PRRS virus HV-NSP2 (500-596) and application thereof

A virus and recombinant bacteria technology, applied in the biological field, can solve the problems of not reaching high fever and insignificant changes in body temperature of sick pigs, and achieve the effect of slowing down the replication ability and reducing the virulence.

Active Publication Date: 2020-06-02
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the body temperature of sick pigs infected with traditional porcine reproductive and respiratory syndrome virus did not change significantly, nor did they reach high fever

Method used

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  • Recombinant PRRS virus HV-NSP2 (500-596) and application thereof
  • Recombinant PRRS virus HV-NSP2 (500-596) and application thereof
  • Recombinant PRRS virus HV-NSP2 (500-596) and application thereof

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Experimental program
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Effect test

Embodiment 1

[0032] Embodiment 1, the acquisition of recombinant PRRS virus HV-NSP2 (500-596), HV-NSP2 (658-777), HV-NSP2 (56)

[0033] 1. Selection of the target segment

[0034] Through the study of the molecular mechanism of PRRS virus-induced hyperthermia, it was found that the non-structural protein 2 (NSP2) of the highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV, referred to as HV) compared with the NSP2 of the classic PRRSV strain VR2332 The amino acid regions 500-596 and 658-777 are the functional regions of highly pathogenic porcine reproductive and respiratory syndrome virus-induced hyperthermia. In order to attenuate the highly pathogenic porcine reproductive and respiratory syndrome virus, it is shown in the 500-596 and 658-777 positions in the non-structural protein 2 (NSP2) of the highly pathogenic porcine reproductive and respiratory syndrome virus HV The amino acid region of the target segment is transformed to construct a recombinant PRRS vi...

Embodiment 2

[0094] Embodiment 2, detection of pathogenicity of recombinant PRRS virus in pigs

[0095] Thirty healthy 4-week-old large white pigs were purchased from the SPF pig farm, and all pigs were further confirmed to be PRRSV negative by PCR. All pigs were randomly divided into 5 groups (6 heads in each group) and the dose of nasal inoculation was 10 5 TCID 50 2mL of virus fluid per mL, 1mL in each nostril. According to the different virus fluid inoculated in each group, they were divided into the following groups:

[0096] Blank control group (MOCK): inoculated with RPMI1640 culture solution;

[0097] Wild-type virus control group: inoculated with wild-type PRRSV highly pathogenic strain HV;

[0098] Recombinant virus experimental group (500-596): inoculated with recombinant PRRS virus HV-NSP2 (500-596);

[0099] Recombinant virus experimental group (658-777): inoculated with recombinant PRRS virus HV-NSP2 (658-777);

[0100] Recombinant virus experimental group (56): inocula...

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Abstract

The invention discloses a recombinant PRRS virus HV-NSP2 (500-596) and application thereof. The cDNA sequence corresponding to the genome of the recombinant PRRS virus is a sequence obtained by deleting DNA molecules shown in the 2833rd-3036th sites of the 5' end of the cDNA sequence corresponding to the genome of a wild type PRRS virus. The invention discloses a molecular mechanism of causing high fever by highly pathogenic PRRSV. Meanwhile, compared with high-pathogenicity PRRSV, the recombinant PRRS virus disclosed by the invention has the advantages that the toxicity is reduced, the replication capacity is also greatly reduced, and a basis is provided for researching and developing a novel attenuated vaccine.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to recombinant PRRS virus HV-NSP2 (500-596) and application thereof. Background technique [0002] Porcine reproductive and respiratory syndrome (PRRS), commonly known as blue ear disease, is one of the important viral diseases that endanger the swine industry all over the world. The cause of porcine reproductive and respiratory syndrome is porcine reproductive and respiratory syndrome virus (porcinereproductive and respiratory syndrome virus, PRRSV). Porcine reproductive and respiratory syndrome virus is a single-stranded positive-sense RNA virus belonging to the order Nidoviridae, family Arteriviridae, and genus Arterivirus. The virus includes two subtypes, the European type (Type I) and the American type (Type II). Currently, the European type is mainly distributed in Europe, while the American type is distributed in North America, South America, and Asia. In 1995, the ...

Claims

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Application Information

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IPC IPC(8): C12N7/01C12N15/40C12N15/63A61K39/12A61P31/14C12R1/93
CPCC12N7/00C12N15/63C07K14/005A61K39/12A61P31/14C12N2770/10021C12N2770/10022C12N2770/10034A61K2039/5254A61K2039/552
Inventor 封文海杜丽唐军王红蕾刘芳魏泽宇
Owner CHINA AGRI UNIV
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