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Preparation method and application of brucella outer membrane vesicle

A technology of outer membrane vesicles and Brucella, which is applied in the field of preparation of Brucella outer membrane vesicles, can solve the problem of low production of Brucella outer membrane vesicles, and achieve effective induction of immune responses and bacterial strains Yield improvement, effect of avoiding carryover

Active Publication Date: 2021-06-25
天康生物制药有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The object of the present invention is to provide a kind of preparation method of brucella outer membrane vesicle and application thereof, to solve the technical problem that the output of brucella outer membrane vesicle is not high in the prior art

Method used

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  • Preparation method and application of brucella outer membrane vesicle
  • Preparation method and application of brucella outer membrane vesicle
  • Preparation method and application of brucella outer membrane vesicle

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Experimental program
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Embodiment 1

[0035] The transformation of embodiment 1 Brucella bacterial strain

[0036] The number of OMVs produced by natural strains of Brucella is limited, and the general output does not exceed 0.4 μg / ml of bacterial liquid. Genetic engineering can be used to modify the strains to increase the production of Brucella OMVs.

[0037] The specific method is as follows:

[0038] 1.1 Target gene knockout plasmid construction

[0039] In the present invention, the genes BamD, OMP39, OMP2b, Cgs, and BF3285c related to Brucella cell membrane stability are selected as targets for transformation, and bioinformatics tools are used for comprehensive evaluation to screen out gene sequences that need to be knocked out. Homology arms were designed upstream and downstream of the gene sequence (see Table 1 for primer information), the pIB279 plasmid (including SacB gene) genome was used as a template, the SacB gene was amplified by PCR using SacB-F and SacB-R as primers, and the homology arms were co...

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Abstract

The invention provides a preparation method and application of brucella outer membrane vesicles, and relates to the technical field of biology. The method comprises the step of carrying out transformation by taking one or more of genes OMP39, BamD, OMP2b, Cgs and BF3285c as a target to obtain a gene deletion mutant strain. Through a gene engineering method, potential gene target sites are selected for modification, five gene-deleted strains are successfully obtained, and the result of extracting the outer membrane vesicles through strain culture shows that the yield of the outer membrane vesicles of the modified strains is remarkably improved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a preparation method and application of Brucella outer membrane vesicles. Background technique [0002] Brucellosis is an important zoonotic infectious disease. After infection, people will develop symptoms such as fever and muscle aches, and then gradually lose their labor force. Animal infection mainly causes reproductive disorders. In conclusion, brucellosis can have a major impact on social and economic development. In the past few decades, people have a better understanding of brucellosis through vaccine immunization and prevention and control promotion, but there are still more opportunities to further improve the prevention and control effect and eradicate brucellosis There is work to be done, and the most important thing is to develop safe and effective vaccines. The currently used vaccines are mainly live attenuated vaccines. During the use of vaccines, there is a risk of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/74C12N15/31C12N1/21A61K39/10A61P31/04C12R1/01
CPCC12N15/74C07K14/23A61K39/098A61P31/04Y02A50/30
Inventor 贺笋潘毅平何传雨吴冬玲赵海龙刘梦志李明奇师伟伟
Owner 天康生物制药有限公司
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