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Ubiquitin-mediated recombinant plasmid expressing avian leukosis virus subgroup J gp85, p27 and p10 genes, and construction method and application thereof

A pvax1-ub-gp85-p27-p10, avian leukosis virus technology, applied in the direction of viruses, gene therapy, antiviral agents, etc., can solve problems that have not been reported

Inactive Publication Date: 2016-11-09
GUANGXI UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

At present, there are few reports on ubiquitin-enhanced immune response to poultry pathogens, especially for viral antigens.

Method used

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  • Ubiquitin-mediated recombinant plasmid expressing avian leukosis virus subgroup J gp85, p27 and p10 genes, and construction method and application thereof
  • Ubiquitin-mediated recombinant plasmid expressing avian leukosis virus subgroup J gp85, p27 and p10 genes, and construction method and application thereof
  • Ubiquitin-mediated recombinant plasmid expressing avian leukosis virus subgroup J gp85, p27 and p10 genes, and construction method and application thereof

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Embodiment Construction

[0018] 1.1 Strains

[0019] J subgroup avian leukemia virus strain GX13HG03 (preserved in the inventor's laboratory).

[0020] 1.2 Method

[0021] 1.2.1 Primer design

[0022] Primers were designed according to the gp85, p27, p10 gene sequences of subgroup J avian leukemia virus and chicken ubiquitin gene sequences published in GenBank, and Hind III, BamH I, EcoR I, Xho I and Kpn I were added to different primers Restriction site, all endonucleases were purchased from Bao Biological Engineering (Dalian) Co., Ltd. The sequences of primers for gp85, p27, p10 and ubiquitin gene amplification are shown in Table 1.

[0023] Table 1 Primer sequences

[0024]

[0025] In Table 1, the parts in italics are restriction sites, and the primers were synthesized by Beijing Huada Gene Company.

[0026] 1.2.2 Amplification of gp85, p27, p10, ubiquitin genes

[0027] Amplification of gp85, p27, and p10 genes of GX13HG03 strain: the gp85, p27, and p10 genes were respectively amplified ...

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Abstract

The invention discloses a ubiquitin-mediated recombinant plasmid expressing avian leukosis virus subgroup J gp85, p27 and p10 genes, and the recombinant plasmid consists of avian leukosis virus subgroup J gp85, p27 and p10 genes, chicken ubiquitin Ub gene and eukaryotic expression vector pVAX1. The corresponding construction method is also established and is capable of realizing serial connection of avian leukosis virus subgroup J gp85, p27 and p10 genes and chicken ubiquitin gene and insertion into the vector pVAX1. An in-vitro experiment proves that the recombinant plasmid is successfully expressed after being used to transfect a DF-1 cell. During a chicken immunity experiment, the antibody positive conversion rate of immunized chicken and antibody positive rate of offsprings are both raised. By constructing the ubiquitin-mediated avian leukosis virus subgroup J multigenic co-expression DNA vaccine, the method helps to establish a basis for researching a novel vaccine for avian leukosis virus subgroup J, and provide an effective auxiliary means for purifying avian leukosis virus subgroup J virus.

Description

technical field [0001] The invention belongs to the technical field of J subgroup avian leukemia, and in particular relates to a ubiquitin-mediated recombinant plasmid expressing gp85, p27 and p10 genes of J subgroup avian leukemia virus and its construction method and application. Background technique [0002] Avian leukemia virus subgroup J (ALV-J) is a new subgroup of avian leukemia virus first isolated from white-feathered broiler chickens in 1988. ALV-J, like ALV-A and ALV-B, can cause immunosuppression of the body, cause vaccine immunity failure, and seriously affect the production performance of poultry, resulting in a decline in egg production and egg quality, seriously affecting the health of the poultry industry develop. Compared with ALV-A and ALV-B, ALV-J spreads faster and is more harmful horizontally. At present, the disease has spread to many countries, causing significant economic losses to the poultry industry. With the introduction of ALV-J from white-fea...

Claims

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Application Information

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IPC IPC(8): C12N15/85C12N15/66A61K48/00A61K39/21A61P31/14A61P35/02
CPCC12N15/85A61K39/12A61K48/005A61K2039/53C12N15/66C12N2740/11034
Inventor 韦平王培坤毕玉彧邹广珍杨永立林璐璐秦丽莉彭昊磨美兰韦天超黄腾
Owner GUANGXI UNIV
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