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Method for constructing PRRSV gene deletion vaccine toxin strain by using Nsp2 gene deletion and uses thereof

A PRRSVBJ-4 strain technology, applied in gene therapy, inactivation/attenuation, biochemical equipment and methods, etc., can solve problems such as unproven functions, and achieve the effect of convenient molecular operation

Inactive Publication Date: 2008-07-16
CHINA AGRI UNIV
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  • Abstract
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  • Claims
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Problems solved by technology

However, other than B-cell epitopes, no other functions have been demonstrated

Method used

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  • Method for constructing PRRSV gene deletion vaccine toxin strain by using Nsp2 gene deletion and uses thereof
  • Method for constructing PRRSV gene deletion vaccine toxin strain by using Nsp2 gene deletion and uses thereof
  • Method for constructing PRRSV gene deletion vaccine toxin strain by using Nsp2 gene deletion and uses thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0035] Embodiment 1, reduce PRRSV BJ-4 strain virulence, obtain rescued virus rV63

[0036] Materials used in this example:

[0037] A. Cells and strains

[0038] MARC-145 cells (China Veterinary Drug Administration) were cultured in DMEM medium. PRRSV BJ-4 strain (China Agricultural University) was in 1997 by Yang Hanchun (Yang Hanchun, Guan Shanhong, Yin Xiaomin, et al. Isolation and preliminary identification of porcine reproductive and respiratory syndrome virus. Chinese Journal of Veterinary Medicine, 1997, 23 (10): 9-10) etc. are isolated and preserved, and its whole genome sequencing has been completed (Yang Hanchun et al., 2001 (Yang Hanchun, Huang Fangfang, Guo Xin, et al. Determination and analysis of the whole genome sequence of porcine reproductive and respiratory syndrome virus (PRRSV) BJ-4 strain . Journal of Agricultural Biotechnology, 2001, 9(3): 212-218) (GenBank accession number is AF331831).

[0039] B. Vector, bacterial strain and PRRSV BJ-4cDNA recombin...

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Abstract

The invention discloses a method for utilizing Nsp2 gene deletion for constructing a PRRSV gene deletion vaccine virus strain and the application thereof. The method for lowering the toxicity of the PRRSV BJ-4 strain of the invention is that a nucleotide segment with the length of 69nt in a Nsp2 coding region of the PRRSV BJ-4 strain genome is continuously deleted to obtain the virus strain with reduced toxicity; the nucleotide sequence of the 69nt nucleotide segment is shown as the 2795 to 2863 sites from the 5' tail end of GenBank Accession Number AF331831. The Nsp2 of the virus rV63 which is prepared by the method for lowering the toxicity of the PRRSV BJ-4 strain deletes 21 amino acids, which has the potential to develop a marker vaccine and can identify the diagnosis method of the deleted polypeptide. When in the development process of vaccine, the invention can utilizes a reverse genetic technical platform which is already established for carrying out the transformation of GP5 glycosylation sites, thus overcoming the suppression factors which affect the neutralizing epitope and further improving the level of the vaccine-induced neutralizing antibody.

Description

technical field [0001] The invention relates to a method for constructing a PRRSV gene deletion vaccine strain by using Nsp2 gene deletion and its application. Background technique [0002] Porcine reproductive and respiratory syndrome (Porcine reproductive and respiratory syndrome, PRRS) was originally called "porcine mystery disease". One of the most important diseases of the industrial economy. The main features of the disease are reproductive failure in breeding pigs, increased mortality in suckling piglets, respiratory disease in pigs of all ages, especially in suckling and nursery pigs, and influenza-like symptoms in growing and finishing pigs. [0003] As the causative agent of PRRS, porcine reproductive and respiratory syndrome virus (Porcine reproductive and respiratory syndrome virus, PRRSV) was first isolated in the Netherlands and the United States, and served as the prototype virus of the European type (LV) and the American type (VR-2332) respectively. strain....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/04C12N7/01A61K48/00A61P31/14
Inventor 杨汉春郭鑫冉智光陈小云盖新娜
Owner CHINA AGRI UNIV
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