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Fusion protein surface immunogenic protein-target of RNA III activating protein (SIP-TRAP) for preventing bovine mastitis and preparation method and use thereof

A fusion protein and protein technology, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, chemical instruments and methods, etc., can solve the problems of poor immune protection effect, narrow protection range, complex immune procedures, etc., and improve immune protection The effect of reducing the scope, reducing the cost of prevention and treatment, and simplifying the immunization procedure

Inactive Publication Date: 2012-01-04
HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The third object of the present invention is to provide the use of the fusion protein SIP-TRAP, so as to overcome the problems of poor immune protection effect, narrow protection range and complicated immune procedures in the prior art of cow (or sheep) mastitis vaccine

Method used

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  • Fusion protein surface immunogenic protein-target of RNA III activating protein (SIP-TRAP) for preventing bovine mastitis and preparation method and use thereof
  • Fusion protein surface immunogenic protein-target of RNA III activating protein (SIP-TRAP) for preventing bovine mastitis and preparation method and use thereof
  • Fusion protein surface immunogenic protein-target of RNA III activating protein (SIP-TRAP) for preventing bovine mastitis and preparation method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Preparation of SIP-TRAP protein

[0047] 1 material

[0048] 1.1 Strains: Staphylococcus aureus reference strain Newman (Utrecht University, Netherlands), Wood46 (strain preserved in this laboratory) and HLJ855-23-1 strain (strain isolated in this laboratory); Streptococcus agalactiae B03020, 8511 -10 strains (strains isolated from our laboratory). The Staphylococcus aureus Newman strain and Wood46 strain used in this example have been published in the literature (Immunogenicity of Staphylococcus aureus agglutinating factor A. Feng Hao, Zhu Zhanbo, Cui Yudong, etc. Acta Biological Engineering, 2009, 25 (8): 1180-1186); Staphylococcus aureus HLJ855-23-1 strain, Streptococcus agalactiae B03020 strain, and 8511-10 strain have been published in the literature (Immunity Research of Cow Mastitis Multiple Inactivated Vaccine, Zhu Zhanbo, Master Thesis, 2004.5).

[0049] 1.2 Vector and host bacteria: The cloning vector pMD18-T vector was purchased from TaKaRabao Bi...

Embodiment 2

[0082] Example 2 Study on the immune protection effect of SIP-TRAP protein

[0083] 1 material

[0084] 1.1 Bacterial strains and experimental animals: The bacterial strains are the same as in Example 1. The mice used in the immunization experiment were clean-grade Kunming mice purchased from Changchun Institute of Biological Products.

[0085] 1.2 Reagents: expressed and purified SIP-TRAP, Sip, and TRAP proteins; Freund's complete and incomplete adjuvants, DMSO, TEMED, TMB, and Tween-20 are all products of Sigma-Aldrich; goat anti-mouse IgG-HRP is Products from Beijing Zhongshan Jinqiao Biotechnology Company; DAB substrate chromogenic solution was purchased from Wuhan Boster Biological Engineering Co., Ltd.

[0086] 2 Experimental methods

[0087] 2.1 Immunogen preparation Draw equal volumes of purified protein and Freund's adjuvant into two syringes respectively, connect a short section of infusion tube (appropriate length) between the two syringes, tie the connection tig...

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Abstract

The invention provides a fusion protein SIP-TRAP and a preparation method and use thereof. The fusion protein SIP-TRAP is obtained inserting a fusion gene formed by connecting the SIP gene of streptococcus agalactiae and the TRAP gene of staphylococcus aureus into a vector and performing expression and purification. Experiments indicate the protein has high immunogenicity and that the immune effect of the protein is better that that of SIP and TRAP alone and that of the combination of SIP and TRAP. The SIP-TRAP protein provided by the invention can serve as an immunogen to improve the resistance of cow or sheep to infection with streptococcus agalactiae and staphylococcus aureus, simplifies the process of immune operation and has good immune effect.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a fusion protein SIP-TRAP of a SIP protein (Surface Immunogenic Protein) of Streptococcus agalactiae and a TRAP protein (Target of RNA III Activating Protein) of Staphylococcus aureus, and the Protein preparation methods and applications. Background technique [0002] Cow mastitis is an inflammation of the mammary gland tissue of dairy cows, which is caused by many reasons, but it is mainly caused by the infection of pathogenic microorganisms in the mammary gland tissue. It is one of the most important diseases that endanger the dairy industry. There are many types of pathogenic microorganisms that cause mastitis in dairy cows, such as bacteria, fungi, and viruses, and the types of pathogenic microorganisms that are prevalent in different regions are also different. However, statistics show that Staphylococcus aureus (S.aureus) and Streptococcus agalactiae (S.agalactiae) are the two...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62C12N15/70A61K39/116A61K39/09A61K39/085A61P15/14A61P31/04C12R1/46C12R1/445C12R1/19
Inventor 崔玉东朱战波迟佳琦宋佰芬
Owner HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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