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African swine fever virus vaccine and preparation method thereof

An African swine fever virus, African swine fever technology, applied in the field of African swine fever subunit vaccine, can solve problems such as lack of immune protection and poor vaccine safety

Active Publication Date: 2021-06-01
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these vaccines still have poor safety and cannot provide effective immune protection against virulent attacks.

Method used

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  • African swine fever virus vaccine and preparation method thereof
  • African swine fever virus vaccine and preparation method thereof
  • African swine fever virus vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082]Example 1 ASFV P72-Ferritin, ASFV P30-Ferritin, ASFV P54-Ferritin, ASFV P54-Ferritin, ASFV P54-AC-FerritIn Preparation and Effectiveness Detection of Sequence Nanoparticle Vaccine

[0083]I. Preparation and efficacy detection of Asfv p72-ferritin optimized sequence nanoparticle vaccine

[0084]1 Configuration for solutions and medium

[0085]The configuration method for solutions and media refers to the related tools (Joseph et al., The third edition, 2002; Osber, et al, fine molecule biology guide, 1998).

[0086]2 Synthesis of Asfv main crust protein P72 gene sequence and ferritin gene sequence

[0087]The present invention obtains a comparison of 13 ASFV primary casing proteins P72 amino acid sequences from NCBI to obtain a paraqualifier, in order to make the main casin protein P72 isotactic column and iron protein better fusion expressed, using signal peptide analysis software ( Signalp) and transmembrane domain analysis software (TMHMM) analyzed the amino acid sequence of ASFV primary c...

Embodiment 2

[0368]Example 2 PCMV-SHAPQ, PCMV-UBSHAPQ and PCMV-Th1-SHAPQ-TH2 preparation and efficacy detection of viral structure

[0369]First, PCMV-SHAPQ preparation and efficacy detection of viral structure

[0370]1 Configuration for solutions and medium

[0371]The configuration method for solutions and media refers to the related tools (Joseph et al., The third edition, 2002; Osber, et al, fine molecule biology guide, 1998).

[0372]2 Synthesis of the SHAPQ gene sequence.

[0373]The present invention will utilize the sympathy sequences of ASFV phospholin P30, envelope protein P54 and CD2V (hemagglutinin, HA) in Example 1. According to JORD: 2012, the SHAPQ amino acid sequence is designed to design the SHAPQ amino acid sequence, and the ASFV phosphorin P30 amino acid sequence is inserted into the recognition enzyme sequence of the ASFV envelope protein P54 amino acid sequence, and in the ASFV envelope protein P54 amino acid sequence N end Connect the C-terminal of the ASFV CD2V structure protein extract...

Embodiment 3

[0490]Example 3 Construction of recombinant virus in PVL-CAG-Th1-SHAPQ-TH2 baculovirus mammal expression and animal experiment

[0491]1. Construct a PVL-CAG vector

[0492]Specific experimental method reference (Zhang Zhi Fang, Yao Bin, Li Wei, etc.) Expresses the method of exogenous genes in animal cells or animal tissues [P]. China: ZL 201210408558.4.) Conducted A recombinant baculovirus transfer vector of the source gene.

[0493]2. Build a recombinant virus that presents a reporter gene

[0494]2.1 Cloning the gene in a pentastating support vector by cloning the gene in the gene in the gene in the gene in the gene in the gene in the gene in the gene in the gene, the ASFV P54-Ferritin, ASFV P30-Ferritin, ASFV P54-FerritIn, ASFV P30-Ferritin

[0495]ASFV P72-FerritIn, ASFV P30-Ferritin, ASFV P54-Ferritin, ASFV CD2V-AC-FerritIn, and Th1-SHAPQ-TH2 gene fragment enzyme-sized PVLCAG vectors were recovered to the same enzyme digestion. After identification, PVL-CAG-P72-Ferritin, PVL-CAG-P44-Ferritin...

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Abstract

The invention discloses an African swine fever vaccine and a preparation method thereof. According to the invention, the structural proteins P72, P30, P54 or CD2v-AC of the African swine fever virus are respectively displayed on the surface of the cage-shaped structure of the self-assembled ferritin, so that the humoral immune efficacy and width of the vaccine are improved, and the immunogenicity of the structural proteins of the African swine fever virus is improved. Structural proteins P30, P54 and CD2v are recombined to obtain recombinant proteins, and the recombinant proteins are connected with ubiquitin to obtain two recombinant proteins, so that the cellular immune effect of the structural proteins of the African swine fever virus is further improved, and better immune protection is provided for animals. The invention also provides a method for preparing the recombinant protein or the African swine fever vaccine. The African swine fever vaccine provided by the invention can initiate a wide neutralizing anti-African swine fever antibody, not only improves the immune efficacy, but also expands the immune range, provides effective immune protection for virulent infection, and has the potential of becoming a universal safe vaccine with multiple protection effects.

Description

Technical field[0001]The present invention relates to swine fever virus vaccines, and more particularly to African swine fever virus vaccines based on African swine fever self-assembled ferritin nano-antigen, belonging to the field of vaccine in African porcane population.Background technique[0002]African swine fever (ASF) is a kind of strong infectious disease in pigs caused by African swine fever virus (ASFV). The symptoms are mainly high heat, respiratory disorder, severe hemorrhage in all tissue organs, and infected strong drugs. 100%. Since the import of this disease has entered my country in 2018, it has caused a serious blow to China's pig industry. The disease is short, the mortality rate is high, and the pig is huge, and the World Animal Health Organization (OIE) is listed as a legal influence of animal diseases, China is listed as a class of animal diseases. ASFV is a dual DNA virus having a capsule. It has 175 ~ 193KB, with 151 open reading frame ORFS, a total of approxim...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/866A61K39/12A61K39/39A61P31/20
CPCC07K14/005C07K14/195C12N15/86A61K39/12A61K39/39A61P31/20C12N2710/12022C12N2710/12034C07K2319/735C07K2319/00C12N2800/105C12N2800/22C12N2710/14143A61K2039/552A61K2039/55522
Inventor 张志芳李轶女高新桃刘兴健易咏竹张伟业曾振王海宁
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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