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Preparation method and application of rhabdovirus expressed swine fever E2 subunit vaccine

A subunit vaccine and baculovirus technology, which is applied in the field of preparation of swine fever E2 subunit vaccine, can solve the problems of not being able to prevent and control swine fever, and can not distinguish between infected and immunized animals, and achieve good safety and immunogenicity Strong, high-yielding effect

Active Publication Date: 2019-01-11
ZHEJIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, vaccination is still an important measure for the prevention and control of swine fever, but traditional vaccines cannot distinguish between infected and immune animals (Differentiating Infected from Vaccinated Animals, DIVA) due to attenuated vaccines, so they cannot meet the needs of the prevention and control of swine fever

Method used

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  • Preparation method and application of rhabdovirus expressed swine fever E2 subunit vaccine
  • Preparation method and application of rhabdovirus expressed swine fever E2 subunit vaccine
  • Preparation method and application of rhabdovirus expressed swine fever E2 subunit vaccine

Examples

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Embodiment 1

[0049] The expression of embodiment 1 hog fever SP-E2 protein

[0050] 1) Amplification of the target gene

[0051] Primers SP-F, SP-R, E2-F, E2-R were designed according to Genbank, SP signal peptide was amplified by SP-F, SP-R first, and E2 fragment was obtained by amplifying E2-F, E2-R. With SP, E2 as template again, SP-F, E2-R primer amplification obtains target gene SP-E2 (such as figure 2 shown).

[0052] 2) Construction of the shuttle plasmid

[0053] The pFastBacHTB plasmid was digested with EcoRI and XholⅠ restriction endonucleases at 37°C for 2 hours and then recovered. The digested vector and the target gene were recombined with homologous recombination at 37°C for 30 minutes, transformed into Ecoli competent cells, identified by spot picking, and positive bacteria were sent sequencing.

[0054] 3) Acquisition of recombinant bacmid

[0055] Transform the positive plasmid into DH10Bac competent cells, pick white spots after 48h, and carry out PCR identification w...

Embodiment 2

[0066] Example 2 Preparation and animal immunization experiments of the CSF SP-E2 protein subunit vaccine of the present invention.

[0067] The SP-E2 protein expressed in Example 1 was mixed with Seppic 206 water adjuvant at a ratio of 1:1 at 350 rpm for 10 min, sealed and stored at 4°C in the dark.

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Abstract

The invention relates to the technical field of molecular biological and animal biological products and aims to provide a rhabdovirus expressed swine fever E2 subunit vaccine. The vaccine is a signalpeptide-containing swine fever SP-E2 protein expressed by a rhabdovirus expression system, wherein the sequence of signal peptide SP is as shown in SEQ ID NO.1 and the nucleotide sequence of expression E2 protein is as shown in SEQ ID NO.2. After a rat is immunized with the SP-E2 subunit vaccine developed by the invention, a specific antibody for swine fever can be generated more efficiently thanQZ14-E2 and HZ08-E2 proteins, and the comprehensive antibody level is far higher than that of QZ14-E2 and HZ08-E2 proteins; after a pig is immunized, the specific antibody can be induced to generate;the vaccine provided by the invention has high yield, strong immunogenicity and high safety; infected and immunized animals can be distinguished through differential diagnosis; swine fever virus can be fundamentally purified.

Description

technical field [0001] The invention belongs to the technical field of molecular biology and veterinary biological products, and in particular relates to a preparation method and application of a swine fever E2 subunit vaccine expressed by baculovirus. Background technique [0002] Classical swine fever (CSF) is a highly contagious and fatal infectious disease of pigs caused by classical swine fever virus (CSFV). The main features are hemorrhage and splenic infarction, which mainly harm domestic pigs and wild boars, and have caused huge economic losses to the pig industry in the world. The epidemic trend of swine fever has changed a lot, showing the coexistence of typical swine fever and atypical swine fever, the coexistence of recessive infection and persistent infection, and the phenomenon of immune failure occurs from time to time. This new and popular form presents new challenges to the swine industry worldwide. [0003] At present, vaccination is still an important me...

Claims

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Application Information

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IPC IPC(8): C12N15/866C12N5/10C07K14/18A61K39/187A61P31/14
CPCA61K39/12A61P31/14C12N15/86C07K14/005A61K2039/552C12N2710/14043C12N2770/24334Y02A50/30
Inventor 徐慧玲何放方维焕
Owner ZHEJIANG UNIV
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