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A kind of recombinant vaccinia virus vaccine for treating bovine brucellosis and preparation method thereof

A technology of Brucella bovis and vaccinia virus, applied in the directions of biochemical equipment and methods, viruses/phages, antibacterial drugs, etc., to achieve the effect of promoting eradication and purification

Inactive Publication Date: 2011-11-30
河南亚卫动物药业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional live attenuated vaccines still have some shortcomings, and they are still pathogenic bacteria to humans; they can also cause abortion in pregnant animals; adult animals can interfere with serological diagnosis after inoculation, making it difficult to distinguish between infection and immunity
With the development and application of molecular biology and molecular immunology, genetic engineering vaccines have become a research hotspot, but there are no relevant reports on the use of recombinant vaccinia virus vaccines for the prevention and treatment of brucellosis

Method used

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  • A kind of recombinant vaccinia virus vaccine for treating bovine brucellosis and preparation method thereof
  • A kind of recombinant vaccinia virus vaccine for treating bovine brucellosis and preparation method thereof
  • A kind of recombinant vaccinia virus vaccine for treating bovine brucellosis and preparation method thereof

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Embodiment 1

[0024] A method for preparing a recombinant vaccinia virus vaccine, which includes the following steps:

[0025] (One) PCR amplification of bp26 gene

[0026] Using the Brucella bovis S19 genome as a template and bp261 and bp262 as primers, PCR amplifies the Brucella bovis bp26 gene. The primer sequences are as follows:

[0027] Bp261 (upstream primer) 5’-ATGAACACTCGTGCTAGC-3’

[0028] Bp262 (downstream primer) 5’-TTACTTGATTTCAAAAACG-3’

[0029] The reaction system is shown in Table 1 below; the reaction parameters are: pre-denaturation at 96°C for 4min; denaturation at 94°C for 30s, annealing at 65°C for 30s, extension at 72°C for 1min, 30 cycles; extension at 72°C for 10min, and storage at 4°C in a refrigerator.

[0030]

[0031] After the amplification, the PCR products were taken for detection by 1% agarose gel electrophoresis. The electrophoresis buffer is 1×TAE, electrophoresis at 120V constant voltage for 30 minutes, and observe the result of gel electrophoresis with an ultravio...

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Abstract

The invention relates to a recombinant vaccinia virus vaccine for treating brucellosis bovis, which recombines peripheral plasma protein bp26 of Brucella bovis with vaccinia virus to construct the recombinant vaccinia virus. The specific method is as follows: firstly, the Brucella bovis genome is used as a template to amplify the bp26 gene of Brucella bovis by PCR; then the target gene bp26 is cloned into a vaccinia virus vector to construct a recombinant vaccinia virus shuttle plasmid; The plastid transfected the recombinant vaccinia virus shuttle plasmid into BHK-21 cells infected with attenuated vaccinia virus, and screened to obtain the recombinant vaccinia virus of Brucella bovis. The invention recombines the bp26 gene of Brucella bovis with good immunogenicity into the vaccinia virus, which can stimulate the body to resist the infection of various serotypes of brucellosis, and the virulence of the recombinant virus is obviously lower than that of the wild virus strain, providing a new method for the prevention and treatment of brucellosis in my country.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a genetic engineering vaccine, in particular to a recombinant vaccinia virus vaccine for treating bovine brucellosis and a preparation method thereof. Background technique [0002] Brucellosis is a zoonotic infectious disease caused by Brucella, which is widely distributed all over the world. According to reports, more than 170 countries and regions have outbreaks of brucellosis. In animal husbandry, there are bovine brucellosis, sheep brucellosis, swine brucellosis, etc., among which the brucellosis of cattle is the most serious. The bacterium has also been isolated from marine mammals. Historically, the disease was a very serious zoonotic disease in my country. In recent years, the infection rate of Brucella in my country has shown an upward trend, which poses new challenges for the prevention and control of Brucellosis. Existing animal vaccines include live attenua...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/10C12N7/01A61P31/04
Inventor 张遂平李明魁赵全成李贞郭芳茹闫树森辛瑞祥张海涛
Owner 河南亚卫动物药业有限公司
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