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Brucella ghost strain, Brucella ghost vaccine and preparation method

A Brucella and bacteria shadow technology, applied in biochemical equipment and methods, chemical instruments and methods, and microbial-based methods, can solve human and animal infections, inability to distinguish vaccine immunity from wild virus infection, and virulence Residue and other issues

Active Publication Date: 2020-10-30
天康生物制药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the commercialized vaccines mainly used at home and abroad are live attenuated vaccines, which are mainly divided into S19, Rev1, A19, S2, and M5. These vaccines have the advantages of high efficacy and long protection period, but they also have fatal shortcomings, such as poison There is still a risk of infection for personnel and animals, and there is detoxification after vaccination, and it is impossible to distinguish between vaccine immunity and wild virus infection after vaccination
These shortcomings severely limit the role of brucellosis vaccines in the prevention and control of brucellosis

Method used

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  • Brucella ghost strain, Brucella ghost vaccine and preparation method
  • Brucella ghost strain, Brucella ghost vaccine and preparation method
  • Brucella ghost strain, Brucella ghost vaccine and preparation method

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preparation example Construction

[0060] Also provide the preparation method of above-mentioned Brucella ghost bacterial strain according to second aspect of the present invention, described preparation method comprises:

[0061] The plasmid containing the phage cleavage protein E gene and the Staphylococcus aureus nuclease A gene is transformed into the Brucella strain to obtain the Brucella ghost strain.

[0062] The preparation method of the brucella ghost strain provided by the invention has a simple process, and the brucella ghost strain can be obtained through a conventional transfection operation, has relatively low requirements on equipment and experimenters, and is suitable for popularization and application.

[0063] In some preferred embodiments, the plasmid further includes functional elements.

[0064] Connecting functional elements in the plasmid can provide the corresponding functions to the plasmid. For example, when the temperature control element is selected as the functional element of the p...

Embodiment 1

[0084] Preparation and cultivation of embodiment 1 Brucella bacterium ghost strain

[0085] 1. Preparation of Brucella ghost strains

[0086] 1.1 Construction of the Brucella ghost strain is constructed by transforming the plasmid containing the phage lytic protein E gene, the Staphylococcus aureus nuclease A gene (SNA), and the temperature control element into the A19 strain by electroporation.

[0087] 1.2 Phage E gene amplification and cloning

[0088] 1.2.1 E gene amplification

[0089]Using the pUC57-E plasmid as a template, use the E gene amplification primers E-F, E-R for amplification. The reaction conditions are: 94°C pre-denaturation for 5 minutes, 94°C denaturation for 30 seconds, 56°C annealing for 30 seconds, and 72°C extension for 30 seconds. 30 cycles, 72°C extension for 10 minutes. A 50 μl reaction system is used, and the specific components are as follows:

[0090] E gene amplification PCR reaction system table

[0091]

[0092] 1.2.2 E gene cloning

...

Embodiment 2

[0119] The preparation of embodiment 2 Brucella ghost vaccines

[0120] 1. Choose 201 as vaccine adjuvant

[0121] Using the Brucella ghost inactivated antigen provided in Example 1 of the present invention and 201 adjuvant as raw materials, the Brucella ghost vaccine was prepared by emulsification.

[0122] Among them, the antigen content is 300-600 CFU of pre-inactivated Brucella ghost antigen per head, and the mass ratio of antigen and adjuvant is 1:1.

[0123] The emulsification conditions are: temperature 30-34°C, 300-500 rpm, time 10-15min.

[0124] 2. Select 206 as vaccine adjuvant

[0125] Using the Brucella ghost inactivated antigen provided in Example 1 of the present invention and 206 adjuvant as raw materials, the Brucella ghost vaccine was prepared by emulsification.

[0126] Among them, the antigen content is 300-600 CFU of pre-inactivated Brucella ghost antigen per head, and the mass ratio of antigen and adjuvant is 1:1.

[0127] The emulsification condition...

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Abstract

The invention provides a brucella ghost strain, a brucella ghost vaccine and a preparation method, and relates to the technical field of biology. The brucella ghost strain comprises a brucella straincontaining bacteriophage lysate E gene and staphylococcus aureus nuclease A gene. The bacteriophage lysate E gene and the staphylococcus aureus nuclease A gene can express bacteriophage lysate E and staphylococcus aureus nuclease A respectively in a brucella strain, and the brucella antigen can be obtained by the bacteriophage lysate E gene and the staphylococcus aureus nuclease A gene in a biological inactivation mode. Compared with the traditional inactivation mode, the brucella ghost strain reserves the natural structure of the bacterial surface antigenic determinant, and can more effectively stimulate the immune response of the organism after inoculation.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a Brucella ghost strain, a Brucella ghost vaccine and a preparation method. Background technique [0002] Currently, brucellosis is still at the peak of historical epidemics, and the epidemic situation among humans and animals is not optimistic. Animals are the main source of infection, and there is no human vaccine. In recent years, with the increase of breeding density and the increase of regional animal mobility, the epidemic of brucellosis has shown explosive growth, and it is in a historically high epidemic period. Vaccination is the most effective means of disease prevention and control, especially in the process of brucellosis control. Most of the commercialized vaccines mainly used at home and abroad are live attenuated vaccines, which are mainly divided into S19, Rev1, A19, S2, and M5. These vaccines have the advantages of high efficacy and long protection period, but they ...

Claims

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Application Information

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IPC IPC(8): C12N1/21A61K39/10A61P31/04C12R1/01
CPCC07K14/005C12N9/22A61K39/098A61P31/04A61K2039/521C12N2795/00022Y02A50/30
Inventor 贺笋何传雨刘梦志赵海龙任立松吴冬玲李明启李延涛
Owner 天康生物制药有限公司
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