Riemerella anatipestifer DNA vaccine based on RA OmpA gene as well as preparation method and identification method of riemerella anatipestifer DNA vaccine

A technology of Duck Rimer's and DNA vaccines, applied in the biological field, can solve the problem of insufficient immune protection effect of the vaccine, and achieve strong immune protection effect and better immune protection effect

Active Publication Date: 2021-05-28
GUIZHOU INST OF ANIMAL HUSBANDRY & VETERINARY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, the immune protection effect of existing vaccines is not strong enough. Therefore, in order to obtain more recombinant plasmids for eukaryotic expression of the outer membrane protein A gene of Riemerella anatipestifer, the scheme of the present application was obtained after research.

Method used

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  • Riemerella anatipestifer DNA vaccine based on RA OmpA gene as well as preparation method and identification method of riemerella anatipestifer DNA vaccine
  • Riemerella anatipestifer DNA vaccine based on RA OmpA gene as well as preparation method and identification method of riemerella anatipestifer DNA vaccine
  • Riemerella anatipestifer DNA vaccine based on RA OmpA gene as well as preparation method and identification method of riemerella anatipestifer DNA vaccine

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Experimental program
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Effect test

Embodiment 1

[0043] A DNA vaccine based on RA OmpA gene Riemerella anatipestifer, including the eukaryotic expression plasmid pVAX1-OmpA of RA OmpA gene; the eukaryotic expression plasmid pVAX1-OmpA is extracted by Escherichia coli, and carried out with pVAX1 as a carrier Prepared, and each bottle of vaccine contains 100,000 μg of recombinant plasmid PVAX1-OmpA; when inoculated, the neck of ducklings is subcutaneously immunized with 100 μg of recombinant eukaryotic expression plasmid PVAX1-OmpA.

[0044] A preparation method based on RA OmpA gene Riemerella anatipestifer DNA vaccine as described above, comprising the following steps:

[0045] (1) Preparation of polyantiserum: RA OmpA gene was constructed into pET32a+ prokaryotic expression vector, and the recombinant expression plasmid pET32a+-RA-OmpA was transformed into competent cell BL21 to induce expression, and after IPTG induced protein expression, 12% SDS-PAGE After analyzing and confirming the target protein, put the recombinant p...

Embodiment 2

[0056] A DNA vaccine based on RA OmpA gene Riemerella anatipestifer, including the eukaryotic expression plasmid pVAX1-OmpA of RA OmpA gene; the eukaryotic expression plasmid pVAX1-OmpA is extracted by Escherichia coli, and carried out with pVAX1 as a carrier Prepared, and each bottle of vaccine contains 100,000 μg of recombinant plasmid PVAX1-OmpA; when inoculated, the neck of ducklings is subcutaneously immunized with 100 μg of recombinant eukaryotic expression plasmid PVAX1-OmpA.

[0057] The eukaryotic expression plasmid pVAX1-OmpA of the RA OmpA gene was identified by PCR, double enzyme digestion and sequencing. The results showed that the OmpA gene was correctly connected to the PVAX1 vector. The size of the OmpA gene fragment was about 1149bp. The double enzyme digestion and sequencing results were used to identify It is a positive recombinant eukaryotic expression plasmid, named PVAX1-OmpA, and the identification results are as follows: figure 1 , figure 2 shown.

...

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Abstract

The invention discloses a riemerella anatipestifer DNA vaccine based on an RA OmpA gene, wherein the riemerella anatipestifer DNA vaccine comprises an eukaryotic expression plasmid pVAX1-OmpA of the RA OmpA gene; during inoculation, subcutaneous immunization is carried out on the neck of a duckling by using a recombinant eukaryotic expression plasmid PVAX1-OmpA of 100 [mu]g/duckling. The invention also discloses a preparation method and an identification method of the riemerella anatipestifer DNA vaccine based on the RA OmpA gene. The eukaryotic expression plasmid pVAX1-OmpA containing the RA OmpA gene are researched and developed, and the recombinant plasmid PVAX1-OmpA immunizes the duckling so as to induce the generation of a specific immune antibody, so that the strong immune protection effect can be generated, and the recombinant plasmid can be adopted as a candidate strain of the novel DNA vaccine so as to lay the foundation for the further development and research of the novel RA gene engineering vaccine. Compared with the prior art, the riemerella anatipestifer DNA vaccine provided by the invention has a better immune protection effect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a Riemerella anatipestifer DNA vaccine based on RA OmpA gene and its preparation method and identification method. Background technique [0002] Riemerella anatipestifer is a contagious infectious disease caused by Riemerella anatipestifer (RA), also known as duck infectious serositis (infectiousserositis), new duck disease, duck sepsis, Duck plague syndrome, pasteurellosis duck plague, etc. are acute or chronic contact infectious diseases that infect ducks, geese and other poultry. Goose infection was once called goose flu or goose exudative sepsis. have high morbidity and mortality. Riemerella anatipestifer is prevalent in our country and almost spreads in all duck raising areas, causing great losses to the development of the duck industry, especially a serious threat to the booming duck industry in the western region. [0003] At present, scholars at home and abroad ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/02A61P31/04C12N15/79C12N15/31C12Q1/686G01N33/68
CPCA61K39/0208A61P31/04C07K14/195C12N15/79C12Q1/686G01N33/68A61K2039/552A61K2039/54A61K2039/53Y02A50/30
Inventor 徐景峨余波李婷张亚楠
Owner GUIZHOU INST OF ANIMAL HUSBANDRY & VETERINARY
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