Recombinant adenovirus expressing Asian type 1 foot-and-mouth disease virus vlp and its application

A technology for recombinant adenovirus and foot-and-mouth disease virus, applied in the field of recombinant adenovirus, can solve problems such as cloning and assembly difficulties and technical difficulties

Inactive Publication Date: 2012-02-08
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

Because FMDV is a highly mutated virus, the nucleotide sequences of different serotypes such as O-type and Asia1-type foot-and-mouth disease differ by more than 15%. In addition, the genome structure of FMDV is unique, and it is very difficult to clone and assemble correctly. It is technically difficult to express FMD virus-like particles and induce high levels of neutralizing antibodies in animals

Method used

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  • Recombinant adenovirus expressing Asian type 1 foot-and-mouth disease virus vlp and its application
  • Recombinant adenovirus expressing Asian type 1 foot-and-mouth disease virus vlp and its application
  • Recombinant adenovirus expressing Asian type 1 foot-and-mouth disease virus vlp and its application

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Embodiment 1

[0023] Construction, purification and identification of embodiment 1 recombinant adenovirus

[0024] 1. Splicing of Asia1 type FMDV genome P1, 2A gene and 3C gene

[0025] According to the infectious clone of Asia1 type FMDV successfully rescued by the inventor's laboratory (Wang Haiwei, Tu Yabin, Zhou Guohui, etc. Establishment of Asia1 type foot-and-mouth disease virus infectious clone [J]. Chinese Journal of Preventive Veterinary Medicine, 2008, 30 (12): 915 -919) (Yu Li, etc., Asia1 type foot-and-mouth disease virus infectious cDNA and its construction method and application, China Invention Patent Application No. 200810171258.2) Asia1 / YS / CHA / 05 strain genome, respectively design upstream and downstream primers (Table 1) using The P1-2A gene and 3C gene were amplified by RT-PCR method, and the PCR amplification products of pBluescript II SK(+) and P1-2A were digested with KpnI and XhoI respectively, transformed after ligation, and positive clones were screened, and identif...

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Abstract

The invention discloses a recombinant adenovirus expressing Asian type 1 foot-and-mouth disease virus VLP and application thereof. The present invention splices the FMDV capsid protein precursor P1-2A gene and 3C gene together to obtain a fusion gene, and its base sequence is shown in SEQ ID NO:1. The base sequence shown in SEQ ID NO: 1 is connected with the adenovirus shuttle vector to obtain a recombinant adenovirus shuttle vector; the recombinant adenovirus shuttle vector is recombined into human defective adenovirus type 5, screened and identified to obtain a highly efficient The recombinant adenovirus expressing Asian type 1 foot-and-mouth disease virus VLP, its microorganism preservation number is: CGMCC No.3467. The Asian type 1 foot-and-mouth disease virus VLP expressed by the defective adenovirus constructed by the present invention can continuously induce high levels of neutralizing antibodies in mice, and can be used as a vaccine for preventing Asia 1 type foot-and-mouth disease.

Description

technical field [0001] The present invention relates to a recombinant adenovirus, in particular to a recombinant adenovirus expressing Asian type 1 foot-and-mouth disease virus VLP. The present invention also relates to the construction method of the recombinant adenovirus and the application of the recombinant adenovirus in the prevention of foot-and-mouth disease, belonging to foot-and-mouth disease the field of prevention. Background technique [0002] Foot-and-mouth disease (FMD) is an acute, febrile, highly contagious infectious disease of cloven-hoofed animals, and its pathogen is foot-and-mouth disease virus (FMDV). In view of the serious harm caused by foot-and-mouth disease to the world economy, the International Office of Epizootics (OIE) and the United Nations Food and Agriculture Organization (FAO) listed it as the first class A infectious disease, and my country also listed it as the first class of animal infectious diseases. Vaccination is an effective means t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/62C12N7/01A61K39/135A61P31/14C12R1/93
Inventor 于力周国辉王海伟王芳
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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