Brucella vaccine and method for preparing antigen protein used for same

A Brucella and antigenic protein technology, applied in the field of antigenic protein preparation, can solve the problems of strong virulence, difficult cultivation, inability to distinguish vaccine immune antibodies and wild virus infection antibodies, etc., and achieves the realization of quality control, simple cultivation, The effect of high expression

Inactive Publication Date: 2009-12-23
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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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

Method used

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  • Brucella vaccine and method for preparing antigen protein used for same
  • Brucella vaccine and method for preparing antigen protein used for same
  • Brucella vaccine and method for preparing antigen protein used for same

Examples

Experimental program
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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 a brucella vaccine applied to livestock and a method for preparing antigen protein used for same. The antigen protein used for the brucella vaccine is brucella bcsap31 protein, and in particular brucella bcsp31 restructured protein. The method for preparing the restructured protein has steps of: taking bacterium total DNA extracted from the brucella as a template, designing a pair of primers according to ribotide complete sequence of brucella bcsp31 gene to carry out polymerase chain reaction, obtaining bcsp31 protein gene by clone and then transferring the bcsp31 protein gene to a yeast expression vector to obtain a restructured expression vector, importing the obtained restructured expression vector to yeast expression bacterium to obtain restructured pichia pastoris expression bacterium, and carrying out proliferation and induced expression of the restructured pichia pastoris expression bacterium so as to obtain the required antigen protein 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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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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