Brucella abortus immune marking recombinant strain and application thereof in preparation of vaccine

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, easy abortion caused by inoculation of female animals, threat to human and animal health, etc., to promote Effects of Eradication and Purification

Inactive Publication Date: 2010-01-20
CHINA AGRI UNIV
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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

Method used

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  • Brucella abortus immune marking recombinant strain and application thereof in preparation of vaccine
  • Brucella abortus immune marking recombinant strain and application thereof in preparation of vaccine
  • Brucella abortus immune marking recombinant strain and application thereof in preparation of vaccine

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Embodiment 1

[0040] Embodiment 1, the construction of inactivated glycosyltransferase gene recombinant strain

[0041]According to the glycosyltransferase gene in the genome of Brucella abortus (GenBank AccessionNumber: 3787661), two pairs of primers were designed, the first pair of primers was used to amplify the homology arm DA5-1, and the second pair of primers was used to amplify DA5 -2. A third pair of primers was designed to amplify the kan resistance gene fragment in the pKD4 plasmid. Homology arm DA5-1 and homology arm DA5-2 are connected to form DNA fragment DA5; insert DA5 into pEASY-T1Simple vector to obtain recombinant vector pEASY-T1-DA5; insert kan resistance gene fragment into pEASY-T1-DA5 Between DA5-1 and DA5-2, the recombinant vector pEASY-T1-KDA5 was obtained; the KDA5 fragment obtained by digestion of pEASY-T1-KDA5 was inserted into the pBluescripts II KS vector, and finally the recombinant vector pBluescripts IIKS-KDA5 was obtained. Homologous recombination between r...

Embodiment 2

[0097] Embodiment 2, identification of recombinant strain RBA5 of Brucella abortus immunolabeled by inactivating glycosyltransferase gene

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Abstract

The invention discloses brucella abortus immune marking recombinant strain and the application thereof in preparation of vaccine. The recombinant strain is obtained by inactivating glycosyltransferase gene in the brucella abortus. The recombinant bacterial strain has weaker toxicity, can be taken as vaccine to immunize animals without inactivation, and can distinguish immune vaccinated animals from clinically diseased animals by means of a serological technology. By being immunized with the strain of the invention, a mouse generates antibody and standard bacterial strain which are different from the existing strain, so that the diseased animals can be distinguished from the immunized animals by the immunological test of animal sera. The invention is applicable to improving the brucella abortus vaccine, researching diagnosis identification agent, and developing the brucella abortus vaccine and the matched diagnosis identification agent, and has significant meaning for eradicating and cleaning the brucella abortus of the animals in the world.

Description

technical field [0001] The invention relates to a Brucella abortus immunomarker recombinant strain and its application in vaccine preparation. 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 th...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/21A61K39/10A61P31/04C12R1/01
Inventor 吴清民何倩倪王真任婕张春燕牛建蕊
Owner CHINA AGRI UNIV
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