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Recombinant pseudorabies virus strain expressing hog cholera virus E2 protein and preparation method and application thereof

A pseudorabies virus and swine fever virus technology, applied in the field of bioengineering, can solve problems such as poor protection effect

Active Publication Date: 2018-11-06
SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The results showed: rPRVJS-2012-E2(gG)-ΔgI / gE vaccine immunization group: Although no test pigs died during the entire monitoring period, the test pigs showed high fever (body temperature 41 ℃ or higher), serum and nasal swabs detected high copies of viral RNA on days 5-7 and days 5-9, respectively, and the protective effect was not good

Method used

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  • Recombinant pseudorabies virus strain expressing hog cholera virus E2 protein and preparation method and application thereof
  • Recombinant pseudorabies virus strain expressing hog cholera virus E2 protein and preparation method and application thereof
  • Recombinant pseudorabies virus strain expressing hog cholera virus E2 protein and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 Construction of recombinant pseudorabies virus vaccine candidate strain expressing classical swine fever virus E2 protein

[0051] Based on the attenuated live vaccine with pseudorabies gene deletion (PRV JS-2012-△gI / gE strain), the gene sequence of the main protective antigen protein (E2 protein) of classical swine fever virus was inserted into Three recombinant pseudorabies virus gene-deleted vaccine strains (rPRV JS- 2012-△gI / gE+E2, rPRVJS-2012-E2(gG)-△gI / gE, rPRV JS-2012-E2(△gI / gE)-△gI / gE). The obtained three strains of recombinant pseudorabies virus were continuously passed on for 20 generations on Vero cells, and the stability of the three strains of recombinant pseudorabies virus was verified by PCR and sequencing, and the expression of exogenous genes of the three strains of recombinant pseudorabies virus was verified by indirect immunofluorescence Condition. The results showed that among the three strains of recombinant pseudorabies virus, except rP...

Embodiment 2

[0076] Example 2 Determination of safety of recombinant pseudorabies virus vaccine candidate strain expressing classical swine fever virus E2 protein

[0077] In order to determine the safety of two recombinant viruses with good genetic stability, rPRV JS-2012-△gI / gE+E2 and rPRV JS-2012-E2(gG)-△gI / gE in Example 1, on piglets, this study respectively The above two strains of recombinant virus were mixed with 10 5.0 TCID 50 The dose per head was used to inoculate 5 negative suckling piglets. The results showed that there was no difference between the two strains of recombinant virus inoculation group and the blank control group, there was no increase in body temperature, no clinical symptoms occurred, and the autopsy observation showed that the tissues and organs were normal. According to the above results, rPRV JS-2012-△gI / gE+E2 and rPRV JS-2012-E2(gG)-△gI / gE are safe to suckling piglets.

[0078] 2.1 Viruses

[0079] Both rPRV JS-2012-△gI / gE+E2 and rPRV JS-2012-E2(gG)-△gI / ...

Embodiment 3

[0087] Example 3 The protective effect of recombinant pseudorabies virus vaccine candidate strains expressing classical swine fever virus E2 protein to virulent strains of classical swine fever virus

[0088]In order to measure the protective effect of rPRV JS-2012-ΔgI / gE+E2, rPRV JS-2012-E2 (gG)-ΔgI / gE two strains of genetic stability good recombinant virus to the strong strain of classical swine fever among the embodiment 1, this After the test pigs were immunized with rPRV JS-2012-ΔgI / gE+E2 and rPRV JS-2012-E2(gG)-ΔgI / gE vaccine candidate strains respectively, they were challenged with shimen strain, a virulent strain of CSFV. The results showed that: after challenged by the virulent strain of classical swine fever virus (shimen strain), the rPRV JS-2012-ΔgI / gE+E2 vaccine immunized group: during the whole monitoring period, the experimental pigs had no obvious body temperature rise and other clinical symptoms, and obtained 100% protection. rPRV JS-2012-E2(gG)-△gI / gE vaccin...

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Abstract

The invention discloses a recombinant pseudorabies virus strain expressing a hog cholera virus E2 protein. An inserted exogenous sequence of the recombinant pseudorabies virus strain includes a CMV promoter, a signal peptide of the hog cholera virus E2 protein and a nucleotide sequence encoding 1-342 amino acids and SV40 polyA of the E2 protein. The invention also discloses a preparation method and application of the recombinant pseudorabies virus low-virulent strain expressing the hog cholera virus E2 protein. The recombinant pseudorabies virus low-virulent strain has the good immune protection effect on pseudorabies virus and hog cholera virus and is suitable as a vaccine candidate strain for prevention and treatment of the pseudorabies and hog cholera.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a recombinant pseudorabies virus strain expressing classical swine fever virus E2 protein and its preparation method and application. Background technique [0002] Classical swine fever is an acute, highly contagious infectious disease caused by swine fever virus. The disease is prevalent worldwide and seriously harmful to the pig industry. It is an important infectious disease for prevention and control and quarantine in various countries. Since the successful development and widespread application of swine fever vaccine (strain C), swine fever has been effectively controlled in my country, and there are few large-scale outbreaks. However, my country has not eliminated swine fever, and in the past ten years, swine fever is still prevalent throughout the country, and the epidemic situation is becoming more and more complicated, which has caused huge economic losses to my c...

Claims

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Application Information

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IPC IPC(8): C12N7/01A61K39/245A61P31/22A61K39/187A61P31/14
CPCA61K39/12A61P31/14A61P31/22C12N7/00C07K14/005A61K2039/70A61K2039/552C12N2710/16734C12N2710/16721C12N2770/24322C12N2770/24334
Inventor 童光志郑浩童武李国新周艳君于海单同领高飞姜一峰虞凌雪
Owner SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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