Recombinant bacterium of brucella melitensis with immunity labeling and use thereof

A technology of Brucella melis and recombinant bacteria, applied in the field of recombinant bacteria of Brucella melis, can solve the problems of high cost, hindering the eradication and purification of brucellosis, and difficult operation, so as to promote the eradication and purifying effect

Inactive Publication Date: 2009-03-18
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). 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, making the diseased animals exist in the natural population for a long time, which seriously threatens human beings and fauna health, impeding the eradication and decontamination of brucellosis in animals

Method used

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  • Recombinant bacterium of brucella melitensis with immunity labeling and use thereof
  • Recombinant bacterium of brucella melitensis with immunity labeling and use thereof
  • Recombinant bacterium of brucella melitensis with immunity labeling and use thereof

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Embodiment 1

[0032] Embodiment 1, the construction of the recombinant bacterium with the Brucella melis of immune labeling

[0033] According to the ATP-binding protein sequence of the O antigen export system in the Brucella gene (GenBank AccessionNumber: BMEI1416), two pairs of primers were designed, one pair of primers was used to amplify the homology arm DM14-1, and the other pair of primers was used to amplify DM14-2. Homology arm DM14-1 and homology arm DM14-2 are connected to form sequence DM14, and sequence DM14 is inserted into pEX18AP plasmid to obtain recombinant vector pEX18AP-DM14. Homologous recombination between the recombinant vector pEX18AP-DM14 and the genome of Brucella melis can obtain the recombinant bacteria of Brucella melis that inactivates the ATP-binding protein coding gene of the O antigen export system.

[0034] 1. Construction of recombinant vector pEX18AP-DM14

[0035] For a schematic diagram of the construction process, see Figure 5 .

[0036] 1. Extracti...

Embodiment 2

[0077] Embodiment 2, the identification of the Brucella recombinant bacterium of inactivation O antigen export system ATP binding protein

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Abstract

The invention discloses a recombinant strain for Brucella melitensis provided with an immune label. The recombinant strain for the Brucella melitensis provided with the immune label is a strain obtained by killing encoding genes of ATP conjugated protein of an O antigen output system in the Brucella melitensis. Compared with an original strain, the obtained strain has lower virulence, and can be used as a vaccine to immunize animals without killing the genes. By utilization of the strain to immunize a mouse, immunoreaction states different from those of the standard strain and the prior vaccine strain can be generated in the body of the mouse, and sicken animals can be identified from immune animal groups by immunological detection of serum of the animals. The modification of the brucelliasis vaccine and the research of a diagnosis and identification agent have important significance in developing a brucelliasis gene deletion marker vaccine and a matched diagnosis and identification agent and promoting the elimination and purification of brucelliasis of the animals all over the world.

Description

technical field [0001] The invention relates to a recombinant bacterium of Brucella melis with immune marker 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). 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, making the diseased animals exist in the natural population for a long time, which seriously threatens human beings and the health of the fauna, hinde...

Claims

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Application Information

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IPC IPC(8): C12N1/21A61K39/10A61P31/04C12R1/01
Inventor吴清民牛建蕊刘文娟任婕李娜张兴林杨羽刘文晓王真张春燕
OwnerCHINA AGRI UNIV