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Optimized porcine circovivus type 2 recombinant adenovirus construction method

A technology of recombinant adenovirus and porcine circovirus, applied in chemical instruments and methods, biochemical equipment and methods, viruses, etc., can solve the problems of lack of vaccines and affecting immune response, achieve easy concentration and storage, and improve immunogenicity Sexuality and low pathogenicity

Inactive Publication Date: 2017-02-15
NORTHWEST A & F UNIV
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Problems solved by technology

[0007] Some scholars have conducted relevant research on PCV2 recombinant adenovirus vector vaccine, but there is no corresponding commercial vaccine. The reason is that the immune response caused by the protein of the adenovirus vector itself will seriously affect the body's immune response to PCV2Cap

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  • Optimized porcine circovivus type 2 recombinant adenovirus construction method
  • Optimized porcine circovivus type 2 recombinant adenovirus construction method
  • Optimized porcine circovivus type 2 recombinant adenovirus construction method

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Embodiment Construction

[0034] The present invention will be further described below in conjunction with the embodiments and with reference to the accompanying drawings.

[0035] An optimized construction method of porcine circovirus type 2 recombinant adenovirus, in which Intron A and WPRE are added to the adenovirus shuttle vector to complete the construction of the recombinant adenovirus.

[0036] Specifically, the construction of the recombinant adenovirus specifically includes the following steps:

[0037] S1 sequentially connected the genes encoding Intron A, Cap and WPRE to the vector pUC57, sequenced and identified it, and named it pUC-Intron A-Cap-WPRE;

[0038] S2 connects IntronA, Cap and WPRE to pShuttle-CMV. Digest pUC-Intron A-Cap-WPRE and pShuttle-CMV with Xho I and EcoR V, then connect Intron A-Cap-WPRE and pShuttle-CMV, transform DH5a, pick bacteria, shake bacteria, extract plasmids, PCR identification and After the identification of single and double enzyme digestion is correct, i...

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Abstract

The invention relates to an optimized porcine circovivus type 2 recombinant adenovirus construction method. The construction of recombinant adenovirus is completed by cloning Human cytomegalovirus first intron (Intron A) and woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) into an adenovirus shuttle vector. The construction method provided by the invention has the advantages that a Cap expression quantity is improved, so that adenovirus usage dose and adenovirus protein autoimmune response can be relieved and the preparation efficiency of the porcine circovivus type 2 recombinant adenovirus can be improved.

Description

technical field [0001] The invention relates to the technical field of animal virus genetic engineering, in particular to a method for constructing an optimized porcine circovirus type 2 recombinant adenovirus. Background technique [0002] Adenovirus has the advantages of strong susceptibility, low pathogenicity, wide host range, good stability, high virus titer, easy concentration and storage, high efficiency of mediating gene transfer, and ability to accommodate large gene fragments. The background has been studied quite clearly, so adenovirus vector has been widely studied and applied in vaccine preparation, and is one of the most promising vectors, but adenovirus itself has some shortcomings that limit its application. [0003] As a vaccine carrier, the biggest problem of adenovirus vector is that the body’s immune response against the vector is too strong. Now there are several strategies to solve this problem. These strategies include: 1. Chemical method, that is, che...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/861
CPCC07K14/005C12N15/86C12N2710/10043C12N2750/10022C12N2830/42C12N2830/48C12N2830/60
Inventor 童德文黄勇李德龙赵晓民
Owner NORTHWEST A & F UNIV
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