Small RNA (ribonucleic acid) related to virulence of Brucella and application of small RNA in preparation of weak virulence Brucella

A Brucella, virulence technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, medical preparations containing active ingredients, etc., can solve the problem of less research and achieve the effect of understanding the pathogenic mechanism

Active Publication Date: 2019-04-09
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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Problems solved by technology

According to speculation, Brucella should theoretically have 100 to 200 sRNAs. Currentl...

Method used

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  • Small RNA (ribonucleic acid) related to virulence of Brucella and application of small RNA in preparation of weak virulence Brucella
  • Small RNA (ribonucleic acid) related to virulence of Brucella and application of small RNA in preparation of weak virulence Brucella
  • Small RNA (ribonucleic acid) related to virulence of Brucella and application of small RNA in preparation of weak virulence Brucella

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Embodiment 1

[0046] 1 Materials and methods

[0047] 1.1 Strains, plasmids and vectors

[0048] Brucella M28 is preserved by the biosafety level 3 laboratory of Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences. Plasmids pSP72 and pBlue-Kan (recorded in "Construction of ERG6 gene deletion mutant strain of S. Period, Lei Jie et al., which are preserved and provided by the Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences.) are preserved by this laboratory. DH5α competent cells were purchased from Nanjing Nuoweizan. 6-7-week-old SPF grade BALB / c mice were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.

[0049] All experiments involving Brucella strains and their pathogenicity were performed in biosafety level 3 laboratories.

[0050] 1.2 Reagents

[0051] Brucella Selective Supplement was purchased from OXOID, and TSB and TSA were purchased from BD Company. Super-Fidelity DNA Polymerase, Mu...

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Abstract

The invention discloses a small RNA (ribonucleic acid) related to virulence of Brucella and application of the small RNA in preparation of the weak virulence Brucella. The application Northern blot proves an sRNA Clu7 exists in a virulent Brucella strain M28, a sRNA defective strain M28 delta Clu7 is constructed by adopting a homologous recombination method, and the affection of the sRNA on the virulence of the M28 is evaluated by taking a mouse macrophage RAW264.7 and a Balb/c mouse as models. A result shows that the replication capability and the survival capability of the virulent strain M28 in the mouse macrophage RAW264.7 and the mouse can be remarkably reduced after deficiency of the sRNA. When in a fifth week of inoculation, the M28 delta Clu7 is completely eliminated from a body ofthe mouse, and the virulence of the M28 delta Clu7 is remarkably lowered in comparison with the M28. A mouse virulence attacking protection experiment shows that the protection effect that the mouseresists the infection of the virulent strain M28 by the immune M28 delta Clu7 can reach the protection force of a vaccine strain M5-90. The results indicate that the sRNA Clu7 has a significant decisive effect on the virulence of the Brucella. The small RNA has a significant promoting effect on a mechanism of disclosing the virulence of the Brucella. According to the small RNA, a novel technical means is provided for research and development of a novel candidate vaccine strain of the Brucella.

Description

technical field [0001] The invention relates to a small RNA related to the virulence of Brucella, and also relates to the application of the small RNA in preparing attenuated Brucella. The invention belongs to the field of biotechnology. Background technique [0002] Brucellosis (brucellosis), referred to as brucellosis, is a zoonotic chronic bacterial infectious disease caused by Brucella. It is widely distributed all over the world. my country is also a serious epidemic area of ​​brucellosis. The number of new cases of human per year ranks first in the world. Brucellosis has caused great damage to the health of our people and agriculture. People infected with Brucella show symptoms such as undulating fever, endocarditis, and arthritis, and animals infected by it can lead to miscarriage. Unlike enteric pathogens, Brucella virulence does not depend on the expression of specific virulence factors, but on their ability to replicate and survive in host cells. Animal brucello...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N1/21A61K39/10A61P31/04C12R1/01
CPCA61K39/098A61P31/04C12N15/113A61K2039/522C12N2310/10
Inventor 步志高胡森许达
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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