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Recombinant denovirus for expressing foot-and-mouth disease virus type A empty capsid, and application thereof

A technology of recombinant adenovirus and foot-and-mouth disease virus, applied in applications, viral peptides, antiviral agents, etc., can solve the problems of differential diagnosis of difficult-to-infect animals and immunized animals, escape of live virus, etc., and achieve long immune protection period and lasting immunity. Response, high titer effect

Inactive Publication Date: 2012-07-04
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the production of inactivated vaccines requires closed facilities to reproduce virulent viruses, and there is a potential risk of escape of live viruses. Outbreaks of FMD in some parts of the world seem to be related to the residual live viruses in inactivated vaccines
In addition, the antigens used in the preparation of FMD inactivated vaccines are not purified and contain non-structural proteins of the virus, making it difficult to carry out the differential diagnosis of infected animals and vaccinated animals

Method used

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  • Recombinant denovirus for expressing foot-and-mouth disease virus type A empty capsid, and application thereof
  • Recombinant denovirus for expressing foot-and-mouth disease virus type A empty capsid, and application thereof
  • Recombinant denovirus for expressing foot-and-mouth disease virus type A empty capsid, and application thereof

Examples

Experimental program
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Embodiment 1

[0031] Example 1 Construction and identification of recombinant adenovirus

[0032] 1. Synthesis of A-type FMDV subgenome P1, 2A2B and 3C genes

[0033] According to the genome sequence of type A FMDV, the P1-2A2B-3C subgenome (SEQ ID No.1) was synthesized, and the sequenced correct recombinant plasmid was named p-A-P12A3C. Plasmid p-A-P12A3C contains the kozka sequence and start codon ATG and stop codon TAG.

[0034] 2. Construction of recombinant adenovirus shuttle vector

[0035] The p-A-P12A3C plasmid and the adenovirus shuttle vector pShuttle-CMV were subjected to SalI and EcoRV double enzyme digestion, and the digested products were purified by gel recovery, ligated with the shuttle vector, and transformed into JM109 competent bacteria, which were tested for kanamycin resistance For screening, colonies were picked and inoculated into 5 mL of LB liquid medium containing kanamycin, cultured overnight, plasmids were extracted, and identified by PCR, enzyme digestion and s...

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Abstract

The invention discloses a recombinant denovirus for expressing foot-and-mouth disease virus type A empty capsid, and application thereof. A sub-genome for coding the foot-and-mouth disease virus type A empty capsid, which is shown as SEQIDNo.1, and a denovirus shuttle plasmid are operably connected to form a recombinant denovirus shuttle expression vector; the recombinant denovirus shuttle expression vector and a denovirus backbone vector plasmid co-transform Escherichia coli, and a cloned recombinant denovirus genome existing in a plasmid form is obtained; and the cloned recombinant denovirus genome is linearized and transfects cells, and the replication-deficient recombinant denovirus is obtained. The replication-deficient recombinant denovirus has high toxicity titer, and can form the foot-and-mouth disease virus type A empty capsid in the replication process and stably express foot-and-mouth disease virus empty capsid in the virus propagation process; the expressed foot-and-mouth disease virus empty capsid can constantly induce high-level neutralizing antibody in bodies of mice and resist the attack of the foot-and-mouth disease virus; and the recombinant denovirus can be prepared into a vaccine for controlling foot-and-mouth disease.

Description

technical field [0001] The present invention relates to a recombinant adenovirus, in particular to a recombinant replication-deficient adenovirus expressing an empty capsid of type A foot-and-mouth disease virus and a construction method thereof. The present invention further relates to the application of the recombinant replication-defective adenovirus in preparing a vaccine for preventing or treating foot-and-mouth disease , Belonging to the field of prevention and treatment of foot-and-mouth disease. Background technique [0002] Foot-and-mouth disease is an acute, febrile, highly contagious infectious disease common to pigs, cattle, sheep and other domestic and wild cloven-hoofed animals, with more than 70 susceptible animals. FMD is clinically characterized by vesicles on the oral mucosa, hooves and udder skin. The disease has multiple transmission routes and fast speed, and has repeatedly broken out worldwide, causing huge economic losses. Therefore, the World Organi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/42C07K14/09C12N15/861C12N15/66A61K48/00A61K39/135A61P31/14
Inventor 于力
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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