Recombinant adenovirus rAd-ORF2-TCE and application thereof

一种重组腺病毒、rad-orf2-tce的技术，应用在基因工程领域，能够解决经济损失、不能完全阻止PCV2感染和传播等问题，达到免疫原性加强、提高体液免疫和细胞免疫水平的效果

Inactive Publication Date: 2014-05-21
GUANGDONG WENS DAHUANONG BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, PCV2-associated disease remains one of the most prevalent diseases in the swine industry, causing huge economic losses
Although there are a variety of commercial PCV2 vaccines on the market, these vaccines can effectively control PCV2 infection to a large extent, but PCV2-related diseases are still prevalent, because these vaccines cannot completely prevent PCV2 infection and transmission

Method used

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Embodiment Construction

[0027] The present invention will be described in further detail below in conjunction with the accompanying drawings and specific embodiments.

[0028] 1. Construction of recombinant adenovirus rAd-ORF2-TCE transfer vector

[0029] Referring to the PCV2ORF2 gene sequence published by NCBI and the artificially synthesized TCE gene sequence, the application software Primer5.0 designed the following two pairs of primers, which were synthesized by Shanghai Yingjun Company, and used to amplify the target gene fragment.

[0030] P1 (as shown in SEQ ID NO.2): 5-GA AGATCT ATGACGTATCCAAGGAGGCG-3 is the upstream primer of ORF2, and the underlined part is the BglⅡ restriction site;

[0031] P2 (as shown in SEQ ID NO.3): 5- GCTGCCGCCACCGCCGCTTCCGCCACCGCCGCTTCCACC GCCACC AGGGTTAAGTGGGGGGTCT-3 is the downstream primer of ORF2, and the underlined part is the hydrophobic polypeptide (Gly4Ser) 3linker sequence encoding 15 amino acids;

[0032] P3 (as shown in SEQ ID NO.4): 5- GGTGGCGG...

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Abstract

The invention relates to a recombinant adenovirus rAd-ORF2-TCE. The recombinant adenovirus rAd-ORF2-TCE is obtained through fusing and connecting an immunogenic protein gene ORF2 of PCV2 (Porcine Circovirus Type 2) and three T lymphocyte epitope (TCE) series genes, and then, recombining to an adenovirus vector. The invention further relates to application of the recombinant adenovirus rAd-ORF2-TCE in porcine viral immunization vaccines. According to the recombinant adenovirus rAd-ORF2-TCE disclosed by the invention, an organism can be effectively stimulated to generate a high-level specific antibody and generate T lymphocyte proliferation, and both the concentration of IFN-gamma in blood serum and the concentration of IL-2 in the blood serum are increased remarkably; compared with ORF2 of separate PCV2, the recombinant adenovirus rAd-ORF2-TCE has the advantage that both humoral immunity level and cellular immunity level of an animal body are increased remarkably.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a recombinant adenovirus rAd-ORF2-TCE and its application. Background technique [0002] Porcine circovirus type 2 (PCV2) is a common cause of postweaning multisystemic wasting syndrome (PMWS), porcine dermatitis and nephropathy syndrome (PDNS), congenital tremor and The main pathogenic factors of swine respiratory diseases. PCV2 belongs to the Circoviridae family and is the smallest animal virus found so far. PCV2 has no envelope, is icosahedral, and has a diameter of about 17 nm. The virus contains 11 open reading frames, of which ORF1 and ORF2 are the two largest coding frames. ORF1 encodes viral replication proteins Rep and Rep, which are closely related to viral replication. ORF2 encodes the main structural protein of the virus, which contains virus-specific epitopes and can induce protective immunity against PCV2 infection. Therefore, capsid protein (capsid protein, Ca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01A61K48/00A61K39/125A61P31/20C12R1/93
Inventor 罗满林陈瑞爱莫永正高娟
Owner GUANGDONG WENS DAHUANONG BIOTECH
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