Trans-membrane form porcine circovirus type 2 recombinant adenovirus and construction method thereof
A technology of recombinant adenovirus and porcine circovirus, which can be used in antiviral agents, virus antigen components, recombinant DNA technology, etc., can solve the problems of economic losses in the pig industry and the lack of a corresponding commercial PCV2 recombinant adenovirus vector vaccine. , to achieve the effect of enhancing immunogenicity
Inactive Publication Date: 2018-01-12
NORTHWEST A & F UNIV
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Problems solved by technology
The first outbreak of the disease in Canada in 1991, followed by reports of the disease in many countries, PCVAD has caused huge economic losses to the pig industry
PCV2 recombinant adenoviral vector vaccines have been studied, such as the unmodified PCV2 recombinant adenoviral vector vaccine Ad-Cap, but there is no corresponding commercialized PCV2 recombinant adenoviral vector vaccine used in clinical practice
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[0037] The present invention will be described in further detail below in conjunction with the accompanying drawings and embodiments.
[0038] Adenoviral vectors have become one of the most widely used live viral vectors because of their high transduction efficiency, wide range of host cells, and ability to infect cells at different stages. However, since adenovirus self-proteins (mainly adenovirus penton protein and hexon protein) can induce a strong immune response in the host, thus limiting the application of this vaccine in clinical practice, how to reduce the autoimmune response It has become a key scientific problem that must be solved for the large-scale application of recombinant adenovirus vector vaccines.
[0039] In the present invention, the first intron of human cytomegalovirus (human cytomegalovirus firstintron, Intron A) and woodchuck hepatitis virus post-transcriptional regulatory element (woodchuck hepatitis virus post-transcriptional regulatory element, WPRE)...
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The invention provides a trans-membrane form porcine circovirus type 2 recombinant adenovirus and a construction method thereof. The recombinant adenovirus is prepared by performing adenovirus packaging on a recombinant adenovirus plasmid, wherein the recombinant adenovirus plasmid is formed by homologous recombination of a shuttle vector of the trans-membrane form porcine circovirus type 2 recombinant adenovirus and an adenovirus skeleton vector. Target gene fragment inserted into the recombinant adenovirus shuttle vector comprises Intron A, coexpression secreting type CD40L, trans-membrane form PCV2 Cap and secreting type GMC SF and WPRE. The protein expression is improved, the CD40L, Cap and GMC SF can be co-expressed, the CD40L and GMC SF are secreted outside cells, and the Cap is inlaid onto the cell membrane and then well quickly recognized by antigen-presenting cells. The recombinant adenovirus combines two different action mechanisms, and the vaccine immunogenicity of the adenovirus vector can be enhanced.
Description
technical field [0001] The invention belongs to the field of genetic engineering vaccines, in particular to the construction of transmembrane porcine circovirus type 2 (PCV2) recombinant adenovirus Ad-A-spCD40L-tpCap-spGMCSF-W. Background technique [0002] Adenovirus has the advantages of low pathogenicity, wide host range, good stability, easy recombinant DNA manipulation, high virus titer, easy concentration and storage, and ability to accommodate large gene fragments, and its biological background has been studied quite well. Therefore, adenoviral vectors are widely studied and applied in vaccine preparation, gene therapy of cancer, infectious diseases and genetic diseases, and are considered to be one of the most promising vectors. [0003] As a commonly used vaccine carrier, adenovirus has the biggest problem that its own protein can induce a strong immune response from the host, which limits the application of this vaccine in clinical practice. [0004] One of the me...
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IPC IPC(8): C12N15/861C12N15/66A61K39/12A61P31/20
Inventor 童德文黄勇李德龙
Owner NORTHWEST A & F UNIV