Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Gene-deleted attenuated African swine fever virus strain as well as construction method and application thereof

A technology for African swine fever virus and gene deletion, applied in the field of biological vaccine products, can solve problems such as homologous recombination loss, self-mutation, and virulence reversion

Active Publication Date: 2021-07-16
JINYUBAOLING BIO PHARM CO LTD +2
View PDF4 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the phenomenon that MGF gene fragments such as MGF360-12L, MGF-13L, and MGF-14L are prone to self-mutation and homologous recombination deletion or complementation of the deletion site is very likely to cause the deletion of CD2v using the above-mentioned literature 1. and MGF360 (12L, 13L, 14L) gene vaccine strains to prepare live attenuated vaccines face potential safety risks of virulence reversion
In view of the multifaceted safety problems in the above patent document 1, there is an urgent need for a safer African swine fever virus vaccine product

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Gene-deleted attenuated African swine fever virus strain as well as construction method and application thereof
  • Gene-deleted attenuated African swine fever virus strain as well as construction method and application thereof
  • Gene-deleted attenuated African swine fever virus strain as well as construction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1: Isolation and Identification of African Swine Fever Virus Strains

[0035] 1.1. Virus strains

[0036] The wild strain of African swine fever virus used in the embodiment is the Guangdong strain isolated from the wild by the Poultry Disease Teaching and Research Office of the Veterinary College of South China Agricultural University, named ASFV GZ2018, and the specific operations include the following steps (operated in a biosafety level 3 laboratory ): Weigh 0.1g of the spleen tissue of a dead pig that is detected as positive for ASFV infection, put it into a 2.0mL sterile EP tube, add 1mL of sterile PBS solution into the tube, and then put 3 pieces of sterile tissue for grinding Steel balls, put the EP tube into the tissue grinder, and perform vibration grinding. Note that the grinder is pre-cooled to 4°C in advance and placed in a balanced position. After repeated freezing and thawing of the ground tissue fluid for 3 times, filter it with a 0.45 μm filt...

Embodiment 2

[0040] Example 2: Determination of I177L Gene Deletion Fragment

[0041] Although Manuel V.Borca et al. (Development of a highly effective Africanswine fever virus vaccine by deletion of the I177L gene results in sterile immunity against the current epidemic Eurasia strain, Journal of Virology, 2020; DOI: 10.1128 / JVI.02017-19 Said document 2) It has been proved that the I177L gene segment is deleted in the genome of type II African swine fever virus Georgia2007 / 1 (the Genebank accession number of Georgia 2007 / 1 is NC_044959.1, wherein the I177L gene segment deletion position is No. 174530-174671 (a total of 142bp ) in 112bp; equivalent to 112bp in 174572-174713 in the genome MT496893 of the ASFV GZ2018 strain) can obtain the gene deletion vaccine strain ASFV-G-ΔI177L with good safety and immune protection effect. However, this document 2 does not specifically disclose the position of the deleted I177L gene fragment; and because the gene is very short, the expressed protein wil...

Embodiment 3

[0049] Embodiment 3: Construction of the recombinant virus of deletion MGF360 (12L, 13L, 14L), CD2V and I177L-D gene

[0050] For the knockout of the African swine fever virus virulence gene, it is often necessary to take into account the immune response and protective effect on pigs, as well as the pathogenicity and safety performance. Although the knockout of the virulence gene MGF360 (12L, 13L, 14L) (above document 1), or the knockout of a specific fragment of the I177L gene alone (above document 2) has been proven to be able to obtain attenuated African swine fever virus vaccine strain, and has a good immune protection effect. However, the deletion of multiple virulence genes may cause low virus titers, reduce the immunogenicity of the attenuated virus strain or weaken the protective effect, and even affect the replication ability of the gene-deleted recombinant virus, but cannot rescue the virus. Based on this, the inventors tried to construct a recombinant virus that si...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a gene-deleted attenuated African swine fever virus strain as well as a construction method and application thereof, and belongs to the technical field of biological vaccine products. The gene-deleted attenuated African swine fever virus strain constructed by a homologous recombination method is a gene-deleted strain in which CD2v, MGF (12L, 13L and 14L) and I177L gene segments are deleted at the same time on the basis of an II-type African swine fever virus genome, and compared with a parent strain, the gene-deleted attenuated African swine fever virus strain is obviously attenuated, and stable replication and immunogenicity of the gene-deleted attenuated African swine fever virus strain are not influenced. After an experimental pig is inoculated with the gene-deleted attenuated African swine fever virus strain, the phenomena of obvious body temperature rise, joint swelling, disease attack or death of the experimental pig do not occur, viremia does not occur, and good safety and good immune challenge protection effect are shown. Therefore, the gene-deleted attenuated African swine fever virus strain provided by the invention can be used as a candidate vaccine strain with good safety and immune protection effect.

Description

technical field [0001] The invention belongs to the technical field of biological vaccine products, in particular to a gene-deleted attenuated African swine fever virus strain and its construction method and application, especially to a simultaneous deletion of CD2v, MGF (12L, 13L, 14L) and I177L genes Attenuated African swine fever virus strain of partial fragments, its construction method and application in vaccine preparation. Background technique [0002] African swine fever virus (ASFV) is the only African virus genus in the African virus family, and it is also the only DNA arbovirus genus. The virus is mainly infected by direct contact with the mouth or nasal cavity, and has multiple cyclic transmission modes. It mainly exists in blood, tissue fluid, internal organs, secretions, and excretions. It has the characteristics of high infection rate, rapid spread, and high fatality rate. The pig industry has caused huge economic losses. [0003] The most important method o...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N7/00C12N7/04C12N15/85C12N15/34A61K39/12A61P31/20C12R1/93
CPCC12N7/00C12N15/85C07K14/005A61K39/12A61P31/20C12N2710/12021C12N2710/12022C12N2710/12052C12N2710/12062C12N2710/12034C12N2800/107A61K2039/5254A61K2039/552
Inventor 宋庆庆叶正琴陈鸿军王衡狄栋栋武瑾贤龚浪屠颉谢振华陈坚刘建奇徐丽媛赵丽霞刘英楠张翀宇
Owner JINYUBAOLING BIO PHARM CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com