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Abortus Brucella vaccine strain S19 marked recombinant strain and application thereof

A technology of Brucella abortus and recombinant strains, applied in the direction of bacteria, antibacterial drugs, bacterial antigen components, etc., can solve the problems of strong virulence, difficult operation, hindering the eradication and purification of animal brucellosis, and achieve Low toxicity, promotes eradication and purification effects

Inactive Publication Date: 2009-11-11
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For a long time, the global strategy for the prevention and control of brucellosis has been mainly quarantine-culling (in developed countries) or immunization (in developing countries). The former uses quarantine and culling of sick cattle, sheep, pigs and other animals as a measure, which costs a lot of money , Difficult to operate; the latter can prevent the spread of the disease in the animal population through immunization and auxiliary culling measures, but it is difficult to distinguish the immune animals from the clinically diseased animals, which makes the diseased animals exist in the natural population for a long time, posing a serious threat Human and fauna health, hindering eradication and decontamination of animal brucellosis
[0003] According to the current international and domestic use, the rough vaccine strain RB51 and the vaccine strain S19 are both attenuated live vaccines, but S19 is highly virulent, and it is easy to cause abortion when inoculated into female animals. Serological methods cannot be used to distinguish infection from wild strains; The virulence of RB51 is relatively weak. Although serological differential diagnosis can be made, the immune efficacy of RB51 is controversial and has not been used in my country.

Method used

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  • Abortus Brucella vaccine strain S19 marked recombinant strain and application thereof
  • Abortus Brucella vaccine strain S19 marked recombinant strain and application thereof
  • Abortus Brucella vaccine strain S19 marked recombinant strain and application thereof

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Experimental program
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Effect test

Embodiment 1

[0033] Embodiment 1, the construction of the recombinant bacterial strain of inactivating glucosyltransferase gene

[0034] According to the glucosyltransferase gene (GenBank AccessionNumber: BAbS19_I09300) in the Brucella abortus genome, two pairs of primers were designed, one pair of primers was used to amplify homology arm DA6-1, and the other pair of primers was used to amplify DA6- 2. Homology arm DA6-1 and homology arm DA6-2 are connected to form sequence DA6, and sequence DA6 is inserted into pEX18AP plasmid to obtain recombinant vector pEX18AP-DA6. Homologous recombination between recombinant vector pEX18AP-DA6 and Brucella abortus strain S19 can obtain a recombinant strain with inactivated glucosyltransferase gene.

[0035] 1. Construction of recombinant vector pEX18AP-DA6

[0036] For a schematic diagram of the construction process, see Figure 5 .

[0037] 1. Extraction of Brucella genome

[0038] Take 15 μl of inactivated Brucella abortus strain S19, add 1ml T...

Embodiment 2

[0080] Embodiment 2, the application of recombinant bacterial strain RB6

[0081] Experimental animals: 4-6 weeks old SPF grade Balb / C female mice, purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., license number SCXK (Beijing) 2007-0001.

[0082] 1. Toxicity identification

[0083] The mice were randomly divided into four groups, 30 in each group. The specific vaccination plan is as follows:

[0084] The first group (test group): inoculate the recombinant bacterial strain RB6 that embodiment 1 prepares, every mouse intraperitoneally inoculates dose 0.1ml, contains 10 7 bacteria.

[0085] The second group (control group 1): inoculate Brucella abortus bacterial strain 2308, every mouse intraperitoneally inoculates dose 0.1ml, contains 10 7 bacteria.

[0086] The third group (control group 2): inoculate Brucella abortus bacterial strain S19, every mouse intraperitoneally inoculates dose 0.1ml, contains 10 7 bacteria.

[0087] The fourth group ...

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Abstract

The invention discloses an abortus Brucella vaccine strain S19 marked recombinant strain and an application thereof. The abortus Brucella recombinant strain is a strain obtained by deactivating the glucosyltransferase coding gene. The obtained recombinant strain has small virulence and can be used as vaccines to immunize animals without needs of deactivation. When the recombinant strain is used for immunizing rats, the inoculated animal generates no lipopolysaccharide antibody and the virulent virus strain-infected animal can be distinguished from novel vaccine strain immunized animal by serological reaction. The recombinant strain is beneficial to improving the Brucella vaccine, researching diagnosis and identification reagents and developing the Brucella vaccine and matched diagnosis and identification reagents, and has extremely important significance in deracinating and purifying the global animal Brucella.

Description

technical field [0001] The invention relates to a Brucella abortus vaccine strain S19 marker recombinant bacterium and application thereof. Background technique [0002] Brucellosis is a zoonotic infectious disease, second only to foot-and-mouth disease among large animal infectious diseases, and has important political and economic influences. The epidemic situation in countries around the world is very serious. For a long time, the global strategy for the prevention and control of brucellosis has been mainly quarantine-culling (in developed countries) or immunization (in developing countries). The former uses quarantine and culling of sick cattle, sheep, pigs and other animals as a measure, which costs a lot of money , Difficult to operate; the latter can prevent the spread of the disease in the animal population through immunization and auxiliary culling measures, but it is difficult to distinguish the immune animals from the clinically diseased animals, which makes the d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21A61K39/10A61P31/04C12R1/01
Inventor 吴清民牛建蕊王真任婕杨羽刘文晓刘文娟张春燕
Owner CHINA AGRI UNIV
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