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Classical swine fever virus E2 protein recombinant subunit vaccine taking salmonella flagellin as molecular adjuvant and preparation method of classical swine fever virus E2 protein recombinant subunit vaccine

A subunit vaccine and flagellin technology are applied in the field of swine fever virus E2 protein recombinant subunit vaccine and its preparation, and can solve the problems of inability to produce IgA antibodies and the like

Active Publication Date: 2021-09-14
湖南兀邦生物科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, none of the E2 recombinant subunit vaccines expressed by baculoviruses currently on the market can produce IgA antibodies against CSFV mucosal immune protection.

Method used

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  • Classical swine fever virus E2 protein recombinant subunit vaccine taking salmonella flagellin as molecular adjuvant and preparation method of classical swine fever virus E2 protein recombinant subunit vaccine
  • Classical swine fever virus E2 protein recombinant subunit vaccine taking salmonella flagellin as molecular adjuvant and preparation method of classical swine fever virus E2 protein recombinant subunit vaccine
  • Classical swine fever virus E2 protein recombinant subunit vaccine taking salmonella flagellin as molecular adjuvant and preparation method of classical swine fever virus E2 protein recombinant subunit vaccine

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Embodiment 1

[0033] Example 1 Expression vector construction and protein preparation

[0034]The present invention is based on the E2 protein gene of CSFV 2.1 strain (GenBank accession: JX262391) and the flagellin Flagellum gene of Enterica serovar Typhimurium strain (GenBank accession: CP007581.1), and designs the combination of E2 and Flagellum protein Fusion expression sequence E2-Flagellum (sequence as shown in SEQ ID NO.1, 2373bp in total), wherein E2 has removed the transmembrane region of the C-terminus, and the D3 region of the Flagellum protein is carried out with the (P2) T cell epitope QYIKANSKFIGITEL of tetanus toxin Instead, a rigid Linker (EAAAK) was used between E2 and Flagellum, and a 6×HIS tag was added to the tail of the fusion protein. The final gene sequence was sent to Nanjing GenScript Biotech Co., Ltd. for codon-optimized synthesis.

[0035] PCR amplification primers were designed, and the synthetic E2-Flagellum gene fragment was used as a template to amplify the E2-...

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Abstract

The invention relates to a classical swine fever virus E2 protein recombinant subunit vaccine taking salmonella flagellin as a molecular adjuvant. The classical swine fever virus E2 protein recombinant subunit vaccine comprises a classical swine fever virus E2 protein and a fusion protein of salmonella flagellin with adjuvant effect, and the nucleotide sequence of the gene for coding the fusion protein is as shown in SEQ ID NO.1. After the vaccine prepared from the fusion protein is used for immunizing animals through intramuscular injection and nasal dripping, the vaccine not only can generate specific cellular immunity and humoral immunity, but also can generate high-level antigen specificity IgG and sIgA in serum and mucous membrane secretions correspondingly, and can be used for removing in-vivo CSFV and mucous membrane contact transmission caused by mouth and nose. The vaccine not only can generate immune protection for cells and body fluid of the CSFV, but also can generate mucosal immune protection, can be used for purifying the CSFV, and has a wide application prospect.

Description

technical field [0001] The invention belongs to the field of veterinary vaccines and veterinary biological products, and relates to a genetic engineering vaccine, in particular to a recombinant subunit vaccine of swine fever virus E2 protein with Salmonella flagellin (Flagellum) as a molecular adjuvant and a preparation method thereof. Background technique [0002] Classical swine fever is an acute contagious disease caused by classical swine fever virus (CSFV), which is characterized by fever, diarrhea and intestinal inflammation. It is highly contagious and fatal. Currently endemic areas or countries still adopt attenuated vaccine vaccination as the main means of preventing swine fever. Pigs immunized with attenuated swine fever vaccine cannot be immunologically distinguished from naturally infected wild swine fever virus. Traditional C strain CSFV Live vaccines, despite their stable effects, can no longer meet the needs of China's CSF eradication and purification program....

Claims

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Application Information

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IPC IPC(8): A61K39/12A61K39/39A61P31/14C07K19/00C12N15/85C12N5/10
CPCA61K39/12A61K39/39A61P31/14C07K14/005C07K14/255C12N15/85A61K2039/552A61K2039/575A61K2039/55516A61K2039/55544A61K2039/54C12N2770/24322C07K2319/00C12N2770/24334Y02A50/30
Inventor 余兴龙丁彦彬赵墩喻晓航罗烨罗灵芝郑金刘江鹰
Owner 湖南兀邦生物科技有限公司
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