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A foot-and-mouth disease virus non-structural protein 3b dominant epitope deletion-marked strain and its preparation method and application

A non-structural protein and foot-and-mouth disease virus technology, which is applied to the field of labeling strains with deletion of the dominant epitope of the non-structural protein 3B of foot-and-mouth disease virus and its preparation, can solve the problem of attenuated vaccine virulence regression, affecting vaccine immunogenicity, and large gene fragments. Problems such as virus replication ability

Active Publication Date: 2022-04-12
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the absence of FMDV G-H loop dominant antigenic epitope will seriously affect the immunogenicity of the vaccine, the attenuated vaccine has the risk of virulence reversion, and the deletion of large gene fragments will affect the replication ability of the virus. Therefore, this area still needs to develop more High-quality FMD-labeled virus vaccine candidate strains provide effective technical support for the effective prevention, control and purification of FMD and the construction of a disease-free area in my country

Method used

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  • A foot-and-mouth disease virus non-structural protein 3b dominant epitope deletion-marked strain and its preparation method and application
  • A foot-and-mouth disease virus non-structural protein 3b dominant epitope deletion-marked strain and its preparation method and application
  • A foot-and-mouth disease virus non-structural protein 3b dominant epitope deletion-marked strain and its preparation method and application

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Embodiment 1

[0051] 1. The construction method of the key amino acid deletion or modified full-length clone containing the dominant epitope of FMDV 3B protein:

[0052] The results of alanine scanning showed that the FMDV-specific 3B monoclonal antibody 4B1 recognized the key amino acid peptides of FMDV 3B1, 3B2 and 3B3 proteins as 2 PYAGP 6 . Using the FMD vaccine strain O / ZK / 93-08 infectious clone pOZKF-Z1234 as the backbone, using gene synthesis technology to mutate and modify the amino acids of the dominant epitopes of FMDV non-structural proteins 3B1, 3B2 and 3B3, and try to construct a 3B protein dominant strain FMD-tagged virus with complete depletion of the epitope.

[0053] In Jinweizhi Biological Co., Ltd. gene synthesis containing 3B protein amino acid mutation modification (see figure 1 ) OZK / 93-08 virus Z4 fragment, and cloned into pUC18 vector respectively, and the recombinant plasmids were respectively named pUC-Z43B6. The pUC-Z43B6 recombinant plasmid was digested with ...

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Abstract

The invention provides a foot-and-mouth disease virus non-structural protein 3B dominant epitope deletion-marked strain as well as a preparation method and application thereof, which belong to the technical field of biotechnology and biological products. Foot-and-mouth disease virus non-structural protein 3B dominant epitope deletion marker strain, the foot-and-mouth disease virus obtained by completely missing the dominant epitope on the basis of non-structural protein 3B; GPYAGP to GPAAP mutation, GPYAGP to GPAAP mutation in 3B2, and GPYEGP to GPYEAA mutation in 3B3. The application of the marked virus strain in preparing the foot-and-mouth disease marked vaccine for distinguishing infection and vaccine immunity can meet the purpose of immune prevention and differential diagnosis, and can be used for effective prevention, control and purification of FMD in my country.

Description

technical field [0001] The invention belongs to the technical field of biotechnology and biological products, and in particular relates to a foot-and-mouth disease virus nonstructural protein 3B dominant epitope deletion-marked strain and a preparation method and application thereof. Background technique [0002] Foot and mouth disease (Foot-and-Mouth Disease, FMD) is an acute, febrile, highly contact disease that is caused by Foot-and-Mouth Disease Virus (FMDV) and mainly domestic animals such as pigs, cattle, sheep and wild cloven-hoofed animals. Infectious disease, the disease spreads rapidly, has a high incidence rate, and is extremely harmful, causing great harm to the development of animal husbandry. [0003] Immunization with traditional inactivated vaccines is the most effective means to prevent and control FMD in my country, but conventional inactivated vaccines cannot distinguish between vaccine immunization and wild virus-infected animals, resulting in the inabili...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/01C12N15/86A61K39/135A61P31/14
CPCC12N7/00C12N15/86A61K39/12A61P31/14C12N2770/32121C12N2770/32152C12N2770/32143C12N2800/107C12N2770/32134A61K2039/552
Inventor 李平花刘在新卢曾军马雪青孙普白兴文曹轶梅李坤李冬
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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