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E3 protein used for detection of sheep-pox virus and hyper-immune serum thereof

A technology of sheep pox virus and immune serum, which is applied in the field of sheep pox virus protein and its hyperimmune serum, can solve the problems of high mortality rate and economic loss of sheep raising industry, and achieve the effect of high serum titer

Inactive Publication Date: 2013-09-25
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Sheeppox virus (SPPV) belongs to Poxviridae, Chordatepoxvirinae, and Goatpoxvirus genus. It is mainly transmitted through aerosol, close contact or insect bites. Sheep are the main host of SPPV. All segments can be infected, but the death cases mainly occurred in lambs, and the mortality rate was as high as 100%, which caused significant economic losses to the sheep industry, and was considered to be an economically potential biological weapon.

Method used

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  • E3 protein used for detection of sheep-pox virus and hyper-immune serum thereof
  • E3 protein used for detection of sheep-pox virus and hyper-immune serum thereof
  • E3 protein used for detection of sheep-pox virus and hyper-immune serum thereof

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Embodiment Construction

[0028] Strains and Cells

[0029] Gulang strain of sheep pox virus and Vero cells were preserved by Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences.

[0030] main reagent

[0031] Ex Taq DNA polymerase, pMD18-T simple cloning vector, JM109 competent cells, plasmid extraction kit, xho Ⅰ and Bam HI restriction endonuclease and T4 DNA ligase were purchased from Dalian Bao Biological Engineering Co., Ltd.; gel recovery kit was purchased from AXYGEN Company; pGEX4T-1 expression vector, BL21 (DE3) plysS host bacteria, GST binding resin and nitrocellulose The prime film was purchased from Novagen; agar powder, peptone and yeast powder were purchased from OXOID; ampicillin (AMP), IPTG, Freund’s adjuvant and horseradish peroxidase (HRP)-labeled rabbit anti-goat IgG were purchased from Sigma Company; horseradish peroxidase (HRP)-labeled goat anti-mouse IgG was purchased from Biowrold Company; prestained protein marker and SuperSignal West Femto ch...

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Abstract

The invention discloses an E3 protein used for detection of a sheep-pox virus and a hyper-immune serum, a preparation method and application thereof. The preparation method for the E3 protein comprises the following steps: carrying out PCR amplification by respectively using sequences represented by SEQ ID 1 and SEQ ID 2 as an upstream primer and a downstream primer and genomic DNA of the sheep-pox virus as a template; and connecting an obtained gene with a vector, ect. so as to obtain a recombinant E3 protein. The recombinant E3 protein is used for immunization of a mouse, and a secondary immune blood sample is employed and undergoes separation so as to obtain immune serum. The recombinant E3 protein and the serum provided by the invention can be applied to both detection of the sheep-pox virus and preparation of a detection reagent or kit used for detection of the sheep-pox virus.

Description

technical field [0001] The present invention relates to a protein of sheep pox virus and its hyperimmune serum, and all uses of the protein and hyperimmune serum. Specifically, this patent relates to the E3 protein and its hyperimmune serum used for detection of sheep pox virus, and the use of this protein and its hyperimmune serum. Background technique [0002] Sheep pox is a highly contagious and devastating viral disease among sheep epidemics. It is listed as a legally notifiable animal disease by the World Organization for Animal Health (OIE). Difficulty and death are the hallmarks. Sheep pox is mainly prevalent in some areas of Africa, the Middle East, India, Nepal and Turkey, and it is mainly prevalent in Qinghai, Gansu, Hunan and Inner Mongolia in China. Sheeppox virus (SPPV) belongs to Poxviridae, Chordatepoxvirinae, and Goatpoxvirus genus. It is mainly transmitted through aerosol, close contact or insect bites. Sheep are the main host of SPPV. All segments can be...

Claims

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Application Information

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IPC IPC(8): C12N15/70G01N33/68C07K16/08C07K16/06G01N33/569
Inventor 张强于少雄颜新敏赵志荀吴国华岳华李应国张光培聂福平周晓黎芦晓立李健朱海霞卢昌
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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