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Porcine reproductive and respiratory syndrome virus M protein CTL cell epitopes and application thereof

A technology for respiratory syndrome and pig reproduction, applied in antiviral agents, viral antigen components, viruses/phages, etc., which can solve problems such as insufficient research and lack of comprehensive understanding of immune mechanisms.

Inactive Publication Date: 2010-04-21
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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Problems solved by technology

However, the research on the cellular immunity of this disease is not in-depth, which leads to a lack of comprehensive understanding of the immune mechanism of PRRS

Method used

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  • Porcine reproductive and respiratory syndrome virus M protein CTL cell epitopes and application thereof
  • Porcine reproductive and respiratory syndrome virus M protein CTL cell epitopes and application thereof
  • Porcine reproductive and respiratory syndrome virus M protein CTL cell epitopes and application thereof

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[0038] 1 Materials and methods

[0039] 1.1.1 Experimental materials PRRSV CH-1a strain, PRRSV positive serum, PRRSV negative serum, anti-PRRSVM protein monoclonal antibody were donated by the Pig Disease Laboratory of Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences; WR strain vaccinia virus, recombinant transfer plasmid pSC11, BHK -21 cells, TK - 143 cells and various E.coli competent cells were donated by Researcher Bu Zhigao from the Zoonotic Disease Research Group of our institute.

[0040] 1.1.2 Main reagents pVAX1 plasmid, Lipofectamine TM 2000 Reagent was purchased from Invitrogen Company; pGEX-6p-1 plasmid was purchased from Pharmacia Company. Various restriction endonucleases, ligases, reverse transcriptases, Taq enzymes, Pyrobest enzymes, pMD18-T Vector, and DNA Markers were purchased from TaKaRa Company; plasmid mini-extraction kits and gel recovery mini-kits were purchased from From Shanghai Huashun Biological Reagent Co., Ltd.; ...

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Abstract

The invention discloses porcine reproductive and respiratory syndrome virus M protein CTL cell epitopes and application thereof. The identification of the epitopes comprises the following steps of: fusing and cloning a M protein gene of a porcine reproductive and respiratory syndrome virus CH-1a strain and a mouse ubiquitin gene to form a DNA vaccine and also form recombination viruses, expressing an M protein, of a WR strain vaccinia virus; immunizing a BALB / c mouse according to a Priming-Boosting policy, separating the mouse splenic lymphocyte, culturing and stimulating a CTL short peptide in vitro predicted and synthesized by a bioinformatic method, and using the flow cytometry and enzyme-linked immunospot technology to identify two CTL epitopes that are K93FITSRCRL and F57GYMTFVHF. The identification of the PRRSVM protein CTL cell epitopes lays a certain theoretical foundation for the PRRS cell immune mechanism and the novel epitope pepetide vaccine and has an instructing significance for the theoretical study on PRRS preventing and control technology.

Description

technical field [0001] The present invention relates to the identification and identification of a cell epitope, in particular to the identification and identification of porcine reproductive and respiratory syndrome virus M protein CTL cell epitope, and the invention also relates to a recombinant virus of WR strain vaccinia virus expressing M protein, which belongs to The control field of porcine reproductive and respiratory syndrome. Background technique [0002] Porcine reproductive and respiratory syndrome (Porcine reproductive and respiratory syndrome, PRRS) is a highly contagious disease of pigs caused by porcine reproductive and respiratory syndrome virus (Porcine reproductive and respiratory syndrome virus, PRRSV), also known as "pig blue ear". sick". The disease is characterized by reproductive disorders in pregnant sows (late abortion, stillbirth, mummified fetuses, weak fetuses) and respiratory disease (interstitial pneumonia) in pigs of all ages, especially in p...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/85C12Q1/70A61K39/12A61P31/14C07K7/06G01N33/569C12R1/93
Inventor 吴东来张维军刘霓红白宇林燕童铁钢王群杨涛
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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