Preparation method of Brucella vaccine and antigenic protein for vaccine

An antigen protein, Brucella technology, applied in the direction of bacterial antigen components, microorganism-based methods, biochemical equipment and methods, etc., can solve the virulence reversion, difficulty in cultivation, and inability to distinguish vaccine immune antibodies from wild virus infection antibodies. and other problems to achieve the effect of quality control, simple culture and high expression

Inactive Publication Date: 2011-11-30
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, long-term use of weak 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

Method used

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  • Preparation method of Brucella vaccine and antigenic protein for vaccine
  • Preparation method of Brucella vaccine and antigenic protein for vaccine
  • Preparation method of Brucella vaccine and antigenic protein for vaccine

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Embodiment Construction

[0025] Specific embodiments of the present invention are described as follows:

[0026] The main material used in the present invention

[0027] Bacterial strain: CVCC12 strain of Brucella abortus type 2, purchased from China Veterinary Drug Control Institute.

[0028] Expression strain: GS115 is the yeast strain for expressing the target gene. When used for plasmid transformation, it must be treated with sorbitol to make it competent and become competent cells. This strain can be purchased from Invitrogen.

[0029] pPIC9K vector: a yeast expression vector, purchased from Invitrogen.

[0030] Brucella positive bovine serum: used for qualitative detection of expressed protein, purchased from China Veterinary Drug Control Institute.

[0031] Horseradish peroxidase-labeled rabbit anti-bovine IgG: used for qualitative detection of expressed protein, purchased from Beijing Baierdi Biotechnology Company.

[0032] Primers: All primers for gene amplification and transformation were...

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Abstract

The invention discloses an anti-brucellosis vaccine used by domestic animals and a preparation method of the antigenic protein used in the vaccine. The antigenic protein used in the Brucella vaccine of the present invention is the Brucella bcsp31 protein, especially the Brucella bcsp31 recombinant protein. The preparation method of the recombinant antigenic protein is to use the bacterial total DNA extracted from Brucella as a template, design a pair of primers according to the complete sequence of nucleotides of the Brucella bcsp31 gene, and carry out polymerase chain reaction, clone and obtain the bcsp31 protein gene, Then transfer to the yeast expression vector to obtain the recombinant expression vector, and then introduce it into the yeast expression bacteria to obtain the recombinant Pichia expression bacteria, proliferate and induce the expression of the recombinant Pichia expression bacteria, and obtain the antigen protein used by separation and purification.

Description

technical field [0001] The invention relates to a vaccine against brucellosis used by domestic animals and a preparation method of the antigenic protein used in the vaccine. 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-animals and animals-humans. It is listed as a second-class infectious disease by the country. In recent years, the epidemic situation of brucellosis in my country has rebounded obviously, and has spread to 28 provinces, autonomous regions and municipalities directly under the Central Government; the outbreak of livestock brucellosis has caused huge economic losses to the animal husbandry. The epidemic situation of brucellosis is very serious....

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/10A61P31/04C12N15/81C12P21/02C12R1/84
Inventor 曹小安邱昌庆周继章郑福英蔺国珍宫晓炜
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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