Recombinant bacterium of brucella melitensis and use thereof

A technology of Brucella melis and recombinant bacteria, which is applied in the direction of bacteria, antibacterial drugs, bacterial antigen components, etc., can solve the problem of strong virulence of vaccines, inability to distinguish vaccine strains from diseased animal strains from wild strains, and different genetic backgrounds. Clear and other issues to achieve the effect of promoting eradication and purification

Active Publication Date: 2009-03-18
CHINA AGRI UNIV
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Problems solved by technology

However, the current vaccines are highly virulent and have unclear genetic backgrounds. Although they can prevent the spread of diseases in animals, they cannot distinguish whethe

Method used

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  • Recombinant bacterium of brucella melitensis and use thereof
  • Recombinant bacterium of brucella melitensis and use thereof
  • Recombinant bacterium of brucella melitensis and use thereof

Examples

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Embodiment 1

[0033] Embodiment 1, the construction of Brucella melis recombinant bacteria

[0034] According to the cold shock protein gene sequence (GenBank Accession Number: BMEI0498) in the Brucella gene, two pairs of primers were designed, one pair of primers was used to amplify homology arm DM11-1, and the other pair of primers was used to amplify DM11- 2. Homology arm DM11-1 and homology arm DM11-2 are connected to form sequence DM11, and sequence DM11 is inserted into pEX18AP plasmid to obtain recombinant vector pEX18AP-DM11. Homologous recombination between the recombinant vector pEX18AP-DM11 and the genome of Brucella melis can obtain the recombinant bacteria of Brucella melis with inactivated cold shock protein coding gene.

[0035] 1. Construction of recombinant vector pEX18AP-DM11

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

[0037] 1. Extraction of Brucella genome

[0038] Take 300 μl of inactivated Brucella melis standard strain 16M, add...

Embodiment 2

[0078] Embodiment 2, the identification of the brucella recombinant bacterium of inactivating cold shock protein

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Abstract

The invention discloses a recombinant strain for Brucella melitensis. The recombinant strain for the Brucella melitensis is a bacterial strain obtained by killing encoding genes of cold shock protein in the Brucella melitensis. Compared with an original strain, the obtained strain has virulence lower than that of the prior vaccine M5, and can be used as a vaccine to immunize animals without killing the genes. By utilization of the strain to immunize a mouse, whether a strain separated from the body of a sicken animal is a vaccine strain or a wild strain can be identified by the PCR method. 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 strain of Brucella melis and its application. 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 is immunized and assisted by culling measures. However, the current vaccines are highly virulent and have unclear genetic backgrounds. Although they can prevent the spread of diseases in animals, they cannot distinguish whether the strains isolated from sick animals are vaccine strains or wild strains, making the vaccine strains simple and fast Safety evaluation has become the foc...

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张兴林李娜杨羽刘文晓王真张春燕
Owner CHINA AGRI UNIV
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