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Method of producing brucella vaccine antigen protein

A technique for Brucella abortus and vaccine antigens, applied in the direction of bacterial antigen components, biochemical equipment and methods, chemical instruments and methods, etc., can solve the problems of strong virulence, difficult cultivation, and high requirements for the cultivation of Brucella problems, to achieve the effect of quality control and training

Inactive Publication Date: 2007-08-15
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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Problems solved by technology

[0004] At present, all brucellosis vaccines for animals on the market are attenuated vaccines of different strains. Long-term use of attenuated strains may become more virulent. In addition, the cultivation of Brucella requires high conditions and is difficult to cultivate.
On the other hand, since the current serological detection technology in my country cannot distinguish vaccine immune antibodies from wild virus infection antibodies

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  • Method of producing brucella vaccine antigen protein

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Embodiment Construction

[0022] Specific embodiments of the present invention are described as follows:

[0023] The main material used in the present invention

[0024] Bacterial strain: CVCC12 strain of Brucella abortus type 2, purchased from China Veterinary Drug Control Institute.

[0025] Expression bacteria: BL21 is the Escherichia coli for the expression of the target gene, and CaCl is required for plasmid transformation 2 The treatment makes it competent to become competent cells, and the strain can be purchased from many bioengineering companies.

[0026] pGEX-6P-1 vector: a prokaryotic expression vector, purchased from Promega.

[0027] Brucella positive bovine serum: used for qualitative detection of expressed protein, purchased from China Veterinary Drug Control Institute.

[0028] Horseradish peroxidase-labeled rabbit anti-bovine IgG: used for qualitative detection of expressed protein, purchased from Beijing Baierdi Biotechnology Company.

[0029] Primers: All primers for gene amplif...

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Abstract

The invention discloses a method to prepare antigen protein of abortion brucellosis subunit vaccine and usage, which comprises the following steps: abstracting bacteria total DNA from abortion brucellosis; using the total DNA as mold; adopting property primer; proceeding polyose chain reaction; cloning; getting OMP25 protein gene; transferring into pronucleus expressing carrier; getting restructuring express carrier; leading-in bacillus coli; getting reconstructing large intestine Escherichia; proceeding inducing expression; culturing reconstructing large intestine Escherichia; preparing antigen protein. This protein can be used to prepare abortion brucellosis subunit vaccine.

Description

technical field [0001] The present invention relates to a method for producing the antigenic protein of the Brucella abortus subunit vaccine, specifically a method for preparing the antigenic protein of the Brucella abortus vaccine, especially the production of bovine Brucella abortus by using the expression system of Escherichia coli The invention discloses a method for antigenic protein used in bacterial gene engineering vaccine, and application of the antigenic protein. Background technique [0002] Livestock brucellosis is distributed worldwide. It exists or breaks out in more than 170 countries and regions among more than 200 countries in five continents, especially in many developing countries in South America, Africa and Asia. Brucellosis is mainly infected through the digestive tract, and generally only occurs between animals and animals or between animals and humans. It is listed as a second-class infectious disease by the country. In recent years, the epidemic sit...

Claims

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Application Information

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IPC IPC(8): C12N15/09C12N15/31C12N1/21C07K14/23A61K39/10A61P31/04
Inventor 邱昌庆杨春华曹小安周继章
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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