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Gene associated with toxicity of Brucella and application of gene in evaluation of toxicity of Brucella and preparation of low-toxicity Brucella

A Brucella gene technology, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problems that there are no research reports on Brucella ribosomal protein

Active Publication Date: 2019-04-19
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, there is no relevant research report on Brucella ribosomal protein L31

Method used

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  • Gene associated with toxicity of Brucella and application of gene in evaluation of toxicity of Brucella and preparation of low-toxicity Brucella

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Embodiment 1

[0041] 1 Materials and methods

[0042] 1.1 Strains, plasmids and vectors

[0043] Brucella M28 is preserved by the biosafety level 3 laboratory of Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, plasmid pSP72, pBlue-Kan r (Recorded in "Construction of Saccharomyces cerevisiae ERG6 gene deletion mutant strain, "Journal of Northwest Agricultural Sciences" 2012 No. 03, Lei Jie et al., preserved and provided by Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences.) and pBBR1MCS-4 are both Saved 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. All experiments involving Brucella strains and their pathogenicity were performed in biosafety level 3 laboratories.

[0044] 1.2 Reagents

[0045] TSB and TSA medium were purchased from BD Company. Brucella Selective Supplement was...

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Abstract

The invention discloses a gene associated with the toxicity of Brucella and application of the gene in evaluation of the toxicity of the Brucella and preparation of low-toxicity Brucella. By using a homologous recombination method, a ribosome gene L31 of the Brucella M28 is replaced by a kanamycin gene, and a Brucella L31 gene deletion strain M28deltaL31 is obtained. By using macrophages RAW264.7and a Babl / c mouse model, the influence of the Brucella L31 gene on the toxicity of the Brucella M28 is evaluated. Through results, it is shown that after the L31 gene is deleted, the growth of the M28 cannot be affected, but the duplication survival capacity of the high-toxicity strain M28 in the mouse macrophage system RAW264.7 and the mouse spleen can be obviously reduced, after the L31 gene issupplemented in the M28deltaL31, the toxicity can recover to the level of a wild strain, and it is proved that the gene is associated with the toxicity of the Brucella M28. The gene for predicting the toxicity can be used for research and development of new brucella vaccines, and the gene has a potential application value.

Description

technical field [0001] The invention relates to a novel virulence gene of Brucella, and also relates to the application of the virulence gene in evaluating the virulence of Brucella and preparing attenuated Brucella. The invention belongs to the field of biotechnology. Background technique [0002] Brucella (Brucella) is an important zoonotic bacteria that can infect humans and various animals, and the disease it causes is called brucellosis. After people are infected with brucellosis, symptoms such as arthralgia, muscle pain, wave fever, etc. may occur, and some patients may experience splenomegaly or lymphadenopathy, epididymitis, orchitis, etc. Livestock infection can cause reproductive system diseases such as testicular swelling in male animals and abortion in female animals. The prevalence of the disease is widespread, almost all over the world. At present, my country mainly relies on attenuated live vaccines to prevent the disease, such as sheep breed M5-90, cattle ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31C07K14/23C12Q1/689C12Q1/04C12N1/20C12N15/74C12N15/66
CPCC12N1/20C12N15/66C12N15/74C12Q1/689C07K14/23Y02A50/30
Inventor 步志高胡森许达
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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