Recombinant adeno-associated virus transfer vector containing gD protein gene of variant porcine pseudorabies virus, virus and preparation and application thereof

A porcine pseudorabies virus and porcine pseudorabies technology, applied in the field of genetic engineering, can solve the problems of loss of antigenic determinants, high cost, complex components of inactivated vaccines, etc., and achieve broad market application prospects, convenient operation, and good immune protection Effect

Active Publication Date: 2021-03-02
BRAINVTA (WUHAN) CO LTD +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Classical vaccines have poor protection against mutant strains. It is necessary to use the clinically isolated mutant pseudorabies virus as a gene sample to develop a more effective vaccine to prevent and control pseudorabies. However, the whole virus is used to prepare inactivated vaccines and gene-deleted attenuated vaccines. It is cumbersome and costly, so DNA vaccines, subunit vaccines and live vector vaccines will be the main directions for the development of PRV vaccines
[0006]...

Method used

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  • Recombinant adeno-associated virus transfer vector containing gD protein gene of variant porcine pseudorabies virus, virus and preparation and application thereof
  • Recombinant adeno-associated virus transfer vector containing gD protein gene of variant porcine pseudorabies virus, virus and preparation and application thereof
  • Recombinant adeno-associated virus transfer vector containing gD protein gene of variant porcine pseudorabies virus, virus and preparation and application thereof

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Embodiment 1

[0088] 1. Primer design and PCR amplification

[0089] Using the genome of the wild strain of porcine pseudorabies virus PRV / JXFC / 2015 as a PCR template, the analysis software Oligo6.0 was used to design primers to amplify the gB and gD genes. The amplified fragments were about 2718bp and about 1203bp respectively. The primer sequences are as follows:

[0090] Table 1: PCR Primers

[0091]

[0092] a: Sal I and Spe I restriction sites were designed at both ends. The left end of the primer sequence is the homology arm sequence, the italic part is the Sal I, Spe I restriction site sequence, behind the restriction site is the transcription enhancing kozak sequence, and the underlined part is the gD target gene primer sequence.

[0093]b: XbaI and SalI restriction sites were designed at both ends. The left end of the primer sequence is the homology arm sequence, the italic part is the XbaI, SalI restriction site sequence, the restriction site is followed by the transcription...

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Abstract

The invention belongs to the technical field of gene engineering, and particularly relates to a recombinant adeno-associated virus transfer vector containing a gD protein gene of a variant porcine pseudorabies virus, a recombinant adeno-associated virus, and preparation and application thereof. The recombinant adeno-associated virus (rAAV) is used as a carrier to express the porcine pseudorabies virus gD protein, and the rAAV has the advantages of safety, effectiveness, specificity and the like. The genetically engineered strain prepared by the method expresses the gD immunogenic protein of the epidemic variation porcine pseudorabies virus strain, and has good immunoprotection for multiple epidemic variation porcine pseudorabies virus strains. The mouse is subjected to intramuscular injection of the anterior shin of the right lower limb, then the body can be continuously induced to generate specific neutralizing antibodies and cellular immunity at a high level.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and more specifically relates to a recombinant adeno-associated virus transfer vector containing a mutant porcine pseudorabies virus gD protein gene, a recombinant adeno-associated virus, and its preparation and application. Background technique [0002] Pseudorabies (PR) is caused by pseudorabies virus (PRV). The main symptoms of domestic animals (pigs, cattle, sheep) and wild animals are fever, itching and encephalitis. Since the clinical symptoms are similar to rabies, Therefore, it is also called pseudorabies. PRV has a wide range of hosts in nature, and pigs are the main natural and storage hosts. PRV can infect pigs of different ages and cause different clinical symptoms, especially the most serious damage caused by infection of pregnant sows and suckling piglets. PRV causes reproductive disorders in pregnant sows, neurological symptoms and high mortality in suckling piglets, ...

Claims

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Application Information

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IPC IPC(8): C12N15/864C12N15/66C12N7/01A61K39/245A61P31/22C12R1/93
CPCC12N15/86C12N15/66C07K14/005C12N7/00A61K39/12A61P31/22C12N2710/16722C12N2750/14143C12N2750/14121C12N2750/14152A61K2039/552A61K2039/54C12N2710/16734
Inventor 张胜何晓斌潘杏方文晶冯诚
Owner BRAINVTA (WUHAN) CO LTD
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