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Recombinant adenovirus capable of resisting foot and mouth disease virus replication and infection as well as preparation method and application of recombinant adenovirus

An anti-foot-and-mouth disease virus, recombinant adenovirus technology, applied in the direction of antiviral agents, applications, viruses/phages, etc., can solve the problem of difficult to achieve the protective effect of mutant strains

Inactive Publication Date: 2012-07-18
SOUTH CHINA AGRI UNIV
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  • Abstract
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Problems solved by technology

However, it takes at least seven days for the vaccine to produce a protective effect. At the same time, due to the large number of FMDV serotypes, there is almost no cross-protection between each serotype, so it is difficult for the single-serotype vaccines currently used to achieve protection against different serotypes and subtypes. Mutant strain produces protective effect

Method used

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  • Recombinant adenovirus capable of resisting foot and mouth disease virus replication and infection as well as preparation method and application of recombinant adenovirus
  • Recombinant adenovirus capable of resisting foot and mouth disease virus replication and infection as well as preparation method and application of recombinant adenovirus
  • Recombinant adenovirus capable of resisting foot and mouth disease virus replication and infection as well as preparation method and application of recombinant adenovirus

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Embodiment 1

[0074] The present invention is based on O-type FMDV isolated from different ages and regions, selects highly conserved sequences of FMDV 3D, L, 2B, and VP1 genes to design and synthesize siRNA, clones it into the PSIREN-shuttle shuttle vector, and constructs recombinant plasmids pShuttle-3D, pShuttle- L, pShuttle-2B and pShuttle-VP1, the specific process is as follows (the ligation reaction of the following construction process was operated according to the instructions of T4 DNA ligase, and the positive clone screening was operated according to the molecular cloning experiment guide, and the competent cells used were Escherichia coli DH5α (Bao Biological Engineering Company):

[0075] (1) Dilute the transcription templates of siRNA-3D, siRNA-L, siRNA-2B and siRNA-VP1 to the same concentration (5pmol / μL) with deionized water, respectively, and dilute the complementary The two oligonucleotides (mixed in 5 μL each) were mixed and placed on a PCR amplification instrument, kept a...

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Abstract

The invention discloses a recombinant adenovirus capable of resisting foot and mouth disease virus replication and infection as well as a preparation method and application of the recombinant adenovirus. A transcriptional template of siRNA-3D, a transcriptional template of siRNA-L, a transcriptional template of siRNA-2B and a transcriptional template of siRNA-VP1 are respectively connected with aU6 promoter and cloned into a carrier pAdeno-x to obtain plasmids which are respectively capable of transcribing and have a series connection structure, and then, the plasmids are packaged by packaging cells after being linearized to obtain the recombinant adenovirus rAdeno-3D-L-2B-VP1. The control effect of the recombinant adenovirus rAdeno-3D-L-2B-VP1 is better than the application effect of a recombinant adenovirus rAdeno-2B-VP1. Therefore, a recombinant plasmid provided by the invention can be used for effectively suppressing foot and mouth disease virus replication and infection and resisting foot and mouth disease virus infection.

Description

technical field [0001] The invention belongs to the technical field of foot-and-mouth disease prevention and control, in particular to a recombinant adenovirus resistant to replication and infection of foot-and-mouth disease virus and its preparation and application. Background technique [0002] Foot-and-mouth disease (FMD) is an acute, febrile, highly contagious infectious disease of artiodactyls caused by foot-and-mouth disease virus (FMDV). The infection rate of the disease is high, and the highest incidence rate after infection can reach 100%. There are more than 70 susceptible animals, among which important economic livestock species such as pigs, cattle, and sheep are all susceptible. In view of the fact that FMD not only reduces animal production performance and causes huge direct economic losses, but also affects the international trade of animals and animal products, so the indirect losses caused are even greater, and it will also affect a country's international i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/861A61K48/00A61P31/14
Inventor 陈金顶李银光赵明秋徐艳芳刘文俊代曼曼
Owner SOUTH CHINA AGRI UNIV
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