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African swine fever gene deletion attenuated virus and live vaccine thereof

A technology of African swine fever virus and gene deletion, applied in the field of veterinary biological products, can solve problems such as chronic diseases of vaccines

Pending Publication Date: 2020-11-13
ACAD OF MILITARY SCI PLA CHINA ACAD OF MILITARY MEDICAL SCI INST OF MILITARY VETERINARY MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In recent years, the attenuated vaccine strains that have deleted genes such as ASFV MGF360, MGF505, and EP402R, as well as the natural attenuated strains isolated in nature, have proved to have certain immune effects in pigs, but it was found in field experiments that these natural attenuated viruses and The use of artificial deletion strain vaccines can cause chronic lesions, such as skin ulcers, fever, joint swelling and other adverse reactions (King et al., 2011; Leitao et al., 2001; Revilla et al., 1992)

Method used

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  • African swine fever gene deletion attenuated virus and live vaccine thereof
  • African swine fever gene deletion attenuated virus and live vaccine thereof
  • African swine fever gene deletion attenuated virus and live vaccine thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] L7L gene deletion construct strains ASFV △ L7L

[0039] Cells, and plasmid strains primary porcine alveolar macrophage (PAM) and primary bone marrow macrophages (BMDMs) from 2--3 months old piglets separated PAM cells containing 10% FBS (TBD) of 1640 medium (Gibco) culture, BMDMs required final concentration of 10ng / ml of GM-CSF containing 1640 complete medium with 10% FBS induced culture. African swine fever virus SY-18 Zhu, Genebank accession number: MH766894.1, separated by the Military Veterinary Institute of Epidemiology in 18 years, the strains used in this study is the fourth generation of cell multiplication PAM drugs, in - Aliquot 80 deg.] C. PUC-EGFP expression vector pUC-mCherry and respectively carrying plasmid EGFP green fluorescence and red fluorescent mCherry, and fluorescent genes were expressed under the control of the p72 promoter;

[0040] Method for constructing homologous recombination using homologous recombination vector, a left homology arm to ...

Embodiment 2

[0043] Deletion strain of ASFV △ L8L L8L Gene Construction

[0044] Cells, and plasmid strains primary porcine alveolar macrophage (PAM) and primary bone marrow macrophages (BMDMs) from 2--3 months old piglets separated PAM cells containing 10% FBS (TBD) of 1640 medium (Gibco) culture, BMDMs required final concentration of 10ng / ml of GM-CSF containing 1640 complete medium with 10% FBS induced culture. African swine fever virus SY-18 Zhu, Genebank accession number: MH766894.1, separated by the Military Veterinary Institute of Epidemiology in 18 years, the strains used in this study is the fourth generation of cell multiplication PAM drugs, in - Aliquot 80 deg.] C. PUC-EGFP expression vector pUC-mCherry and respectively carrying plasmid EGFP green fluorescence and red fluorescent mCherry, and fluorescent genes were expressed under the control of the p72 promoter;

[0045] Method for constructing homologous recombination using homologous recombination vector, a left homology arm...

Embodiment 3

[0048] Construction of gene deletion strains L9R of ASFV △ L9R

[0049] Cells, and plasmid strains primary porcine alveolar macrophage (PAM) and primary bone marrow macrophages (BMDMs) from 2--3 months old piglets separated PAM cells containing 10% FBS (TBD) of 1640 medium (Gibco) culture, BMDMs required final concentration of 10ng / ml of GM-CSF containing 1640 complete medium with 10% FBS induced culture. African swine fever virus SY-18 Zhu, Genebank accession number: MH766894.1, separated by the Military Veterinary Institute of Epidemiology in 18 years, the strains used in this study is the fourth generation of cell multiplication PAM drugs, in - Aliquot 80 deg.] C. PUC-EGFP expression vector pUC-mCherry and respectively carrying plasmid EGFP green fluorescence and red fluorescent mCherry, and fluorescent genes were expressed under the control of the p72 promoter;

[0050] Method for constructing homologous recombination using homologous recombination vector, gene to be mis...

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Abstract

The invention discloses an African swine fever gene deletion attenuated virus, further provides an African swine fever gene deletion live vaccine prepared by utilizing the African swine fever gene deletion attenuated virus, and provides a novel attenuated African swine fever virus with gene deletion. By deleting partial or all gene functions of L7L, L8L, L9R, L10L and L11L, compared with a parentstrain, the obtained recombinant strain has the advantages that after a pig is infected, fever and viraemia do not appear or are remarkably relieved, and the test pig is healthy and alive; vaccines prepared from the gene deletion live virus can protect susceptible pigs from ASFV virulent infection or artificial challenge, and can be used for preventing African swine fever.

Description

Technical field [0001] The present invention discloses a gene deletion attenuated African swine fever, the present invention further provides the use of African swine fever gene deletion prepared in the attenuated live vaccines, veterinary biological products belonging to the technical field. Background technique [0002] African swine fever African swine fever virus (ASFV) pigs caused by a highly contagious hemorrhagic disease, the mortality rate approaches 100%, to the pig industry has brought great loss; for the prevention and control of African swine fever in the past mainly taken culling and methods to clear, so far there is no vaccine development success. Full length genome of the African swine fever virus 170-190kb, wherein the constant region is between 125kb, left and right 35kb 15kb variable region. In the presence of a large number of variable region virulence, and immunosuppression, including multigene family of genes, inhibition of apoptosis-related genes, including ...

Claims

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Application Information

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IPC IPC(8): C12N7/00C12N15/85C12N15/34A61K39/12A61K39/295A61P31/20C12R1/93
CPCC12N7/00C12N15/85C07K14/005A61K39/12A61P31/20C12N2710/12021C12N2710/12022C12N2710/12034C12N2710/12052C12N2710/12062C12N2710/12071A61K2039/5254A61K2039/552A61K2039/70
Inventor 扈荣良张静远张艳艳陈腾周鑫韬高玉伟
Owner ACAD OF MILITARY SCI PLA CHINA ACAD OF MILITARY MEDICAL SCI INST OF MILITARY VETERINARY MEDICINE
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