Adenovirus/viruses A replicon chimeric vector hogcholeravaccine and application thereof

A chimeric vector, adenovirus technology, applied in applications, antiviral agents, antibody medical components, etc., can solve problems such as low cell efficiency and low expression efficiency

Active Publication Date: 2012-06-27
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] The key technical problem to be solved by the present invention is to overcome the defects of low cell entry efficiency or low expression efficiency in vivo existing in the existing classical swine fever vector vaccines, and to use adenovirus vectors with high-efficiency delivery capabilities to transport Alphavirus replicon vector to construct a swine fever vaccine based on adenovirus / alphavirus replicon chimeric vector

Method used

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  • Adenovirus/viruses A replicon chimeric vector hogcholeravaccine and application thereof
  • Adenovirus/viruses A replicon chimeric vector hogcholeravaccine and application thereof
  • Adenovirus/viruses A replicon chimeric vector hogcholeravaccine and application thereof

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Embodiment

[0030] 1 Materials and methods

[0031] 1.1 Carriers, cells, biochemical reagents

[0032] The Alphavirus replicon DNA vaccine pSFV1CS-E2 expressing CSFV E2 protein is constructed by this laboratory (this vector is constructed by the inventor's own laboratory, and its construction method is as follows: Based on the Semliki Forest Virus (SFV) replicon, add CMV promoter, SV40 Poly(A) signal and multiple cloning sites). Adenovirus vector system including adenovirus backbone vector pAdEasy-1 and adenovirus shuttle vector pShuttle-CMV and E. coli BJ5183 competent cells were purchased from Stratagene; E. coli JM109 and DH5α competent cells were preserved by our research group. HEK293 cells (purchased from Shanghai Cell Resource Center, Chinese Academy of Sciences) were cultured with DMEM medium (Gibco BRL Company) containing 10% fetal bovine serum and appropriate amount of antibiotics. The non-restriction endonucleases BsiwI, sPeI, PmeI and PacI are all products of NEBBioLabs. Th...

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Abstract

The invention discloses an adenovirus / viruses A replicon chimeric vector hogcholeravaccine and application thereof. In the invention, a viruses A replicon vector expressing a hog cholera virus E2 protein is transmitted by the adenovirus having very strong gene transfer capability, an adenovirus / viruses A replicon chimeric vector vaccine efficiently expressing the CSFV.E2 protein is created, and microbe storage number of the vaccine is CGMCC.No.4146; The chimeric vector vaccine of the invention transmits the viruses A replicon chimeric vector vaccine expressing the CSFV.E2 protein through the adenovirus vector, so that efficient exogenous gene transfer capability of the adenovirus vector is utilized, and advantages of efficient exogenous gene expression of the viruses A replicon vector after self-replication and inducing strong cellular immune response are exerted, so that immunity of the vaccine is greatly enhanced, and immunizing dose is greatly reduced. The chimeric vector vaccine of the invention can induce high-level immune response on a pig body, and can completely protect the pig from attack of fatal hog cholera virulent virus.

Description

technical field [0001] The present invention relates to a swine fever vaccine, in particular to adenovirus / alphavirus replicon chimeric carrier swine fever vaccine, and the invention also relates to the construction method of the swine fever vaccine and its application in the prevention of swine fever, belonging to the control of swine fever field. Background technique [0002] Classical swine fever (CSF) is a severe infectious disease caused by classical swine fever virus (CSFV). notifiable diseases), causing huge economic losses to the pig industry in many countries. CSFV is an enveloped single-stranded positive-strand RNA virus with a genome size of about 12.3kb and only one large open reading frame (ORF). The polyprotein encoded by it is hydrolyzed into four types by the proteases of cells and viruses Structural proteins and 8 nonstructural proteins. Wherein E2 envelope glycoprotein is the main structural protein of CSFV, is the main protective antigen of CSFV, can in...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61K39/187C12N15/40C12N15/861A61P31/14
Inventor 仇华吉孙元李娜李宏宇
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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