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Preparation method of goat pox virus recombinant protein antigen and application thereof

A protein and protein technology, applied in the field of molecular biotechnology and immunity, can solve the problems of immunity and safety defects, vaccines cannot be completely eliminated and controlled, etc., and achieve good antigen reactivity, good antigen immunogenicity, and non-toxic side effects

Active Publication Date: 2019-02-15
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there is no effective treatment for the disease, and only clinical trauma treatment and supportive therapy can be adopted. In terms of prevention and control, goat pox attenuated live vaccine is mainly used for prevention at home and abroad, but this vaccine still cannot completely eliminate and control the disease. There are certain defects in immunity and safety, so it is imperative to create a safe and efficient new vaccine

Method used

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  • Preparation method of goat pox virus recombinant protein antigen and application thereof
  • Preparation method of goat pox virus recombinant protein antigen and application thereof
  • Preparation method of goat pox virus recombinant protein antigen and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Embodiment 1, the preparation of goat pox subunit vaccine

[0068] 1. Preparation of ORF056-C, ORF112 and ORF117-C recombinant proteins

[0069] 1. Cloning of ORF056, ORF117 extracellular region and ORF112 gene and construction of expression vector

[0070] Genomic DNA of goat pox virus GTPV / HBSZ strain (recorded in the following documents: Zhou T, Jia H, Chen G, et al, Phylogenetic analysis of Chinese sheeppox and goatpox virus isolates. Virol J, 2012 9:25), PCR Amplify the extracellular region of ORF056 gene, the extracellular region fragment of ORF117 gene and the full length of ORF112 gene, digest the target fragment and prokaryotic expression vector with restriction endonuclease, connect the target gene and vector with ligase, and transform BL21 competent cells , the expression of the target protein is induced by an inducer, and the expressed protein is purified by a nickel chromatography column. details as follows:

[0071] 1), primer design and PCR amplificati...

Embodiment 2

[0110] Embodiment 2, recombinant protein antigen-specific antibody and neutralizing antibody assay

[0111] 1. Experimental animals and immunity

[0112] Seventy-two female BALB / C mice (18 g) aged 6-8 weeks were purchased from the Experimental Animal Center of Lanzhou University. Divided into 6 groups:

[0113] ORF056 recombinant protein vaccine group (GTPV-ORF056): The ORF056 recombinant protein vaccine in Example 1 was immunized by intramuscular injection into the legs of mice on 0d, 14d and 28d, with a dose of 200 μL (50 μg protein / mouse).

[0114] ORF112 recombinant protein vaccine group (GTPV-ORF112): The ORF112 recombinant protein vaccine prepared in Example 1 was immunized by intramuscular injection into the legs of mice on 0d, 14d and 28d, with a dose of 200 μL (50 μg protein / mouse).

[0115] ORF117 recombinant protein vaccine group (GTPV-ORF117): The ORF117 recombinant protein vaccine prepared in Example 1 was immunized by intramuscular injection into the legs of mi...

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Abstract

The invention discloses a preparation method of a goat pox virus recombinant protein antigen and application thereof. The invention provides a product having the function of 1) or 2): 1) preventing, diagnosing and / or treating animal goat pox, and 2) inhibiting or neutralizing a goat pox virus. The product is any of the following products: protein ORF056, protein ORF112 and protein ORF117; or a composition containing the protein ORF056, protein ORF112 and protein ORF117; or a fusion protein containing the protein ORF056, protein ORF112 and protein ORF117. The experiment of the invention provesthat a mouse immunized by a subunit vaccine prepared by using IMV ORF056, ORF112-encoded protein and EEVORF117-encoded protein can produce a neutralizing antibody for the goat pox virus more than thatproduced by a mouse immunized by a vaccine prepared by single ORF056, ORF112 or ORF117 protein, so that the goat pox virus recombinant protein antigen is an ideal vaccine with the potential goat poxprevention effect.

Description

technical field [0001] The invention belongs to the field of molecular biology technology and immunization technology, and specifically relates to a preparation method and application of a group of goat pox subunit vaccine recombinant protein antigens. Background technique [0002] Goat pox (Goat pox) is a febrile, contact skin and mucous membrane disease caused by goat pox virus of the Poxviridae Chordatepoxvirinae subfamily Goatpoxvirus; in Africa, the Middle East, Central Asia, India and my country It is widely prevalent in Northwest, Northeast and other places, and is listed as one of the animal diseases that must be reported by the World Organization for Animal Health, and it is listed as a first-class animal disease that must be notified in my country. Goatpox is endemic in countries and regions such as Central Asia, India, Central Africa, and North Africa. It is also popular in my country's Heilongjiang, Jilin, Liaoning, Hebei, Gansu, Qinghai, and Inner Mongolia provin...

Claims

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Application Information

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IPC IPC(8): C07K14/065C07K19/00C12N15/39C07K16/08A61K39/275A61P31/20
CPCA61K39/12A61K2039/552A61P31/20C07K14/005C07K16/08C07K2319/00C12N2710/24022C12N2710/24234
Inventor 景志忠陈国华贾怀杰何小兵房永祥牛贤云
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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