Preparation method and application of a candidate outer membrane protein of Aeromonas hydrophila vaccine

Abstract

The invention provides a preparation method and application of a candidate outer membrane protein of Aeromonas hydrophila vaccine, and designs outer membrane protein gene cloning primers; through prokaryotic recombination, using in vitro PCR amplification technology, amplifies target gene fragments, and constructs pET32a prokaryotic Express the recombinant plasmid of the vector, transform the recombinant plasmid to BL21 competent, and obtain a high-expression strain; induce the high expression of OMP P5 protein by IPTG inducer and purify the expressed protein by Ni-NTA resin column; the purified outer membrane protein and Freund's Insulin The complete adjuvant is completely emulsified at a volume ratio of 1:1, and the obtained sample is used as the vaccine protein of the Aeromonas hydrophila vaccine. In the present invention, the Aeromonas hydrophila subunit vaccine (outer membrane protein) can be used to obtain an immune protection rate of more than 90%. Compared with traditional vaccines, the subunit vaccine has low toxic and side effects, strong immunogenicity and long duration, High antibody titer and other advantages.

Description

technical field [0001] The invention relates to the fields of genetic engineering and immunology, in particular to the preparation and application of a candidate outer membrane protein of Aeromonas hydrophila vaccine. Background technique [0002] Aeromonas hydrophila ( Aeromonas hydrophila ) is a Gram-negative brevibacterium, a common opportunistic pathogen widely present in the aquatic environment, and is a typical zoonotic pathogen of humans, animals, and fish. The extensive attention of the academic, veterinary and medical circles has become the object of public health attention. As a large breeding country, Aeromonas hydrophila seriously damages the development of my country's breeding industry, and even threatens human safety. At present, antibiotics are the main therapeutic drugs against aquatic pathogens such as Aeromonas hydrophila. However, long-term and large-scale use of antibiotics is likely to produce drug-resistant strains, and even super-strong bacteria. Ef...

AI Technical Summary

Problems solved by technology

At present, researchers (Sun Li et al. 2013; Zheng Zonglin et al. 2015) have found that the immune protection rate is 60-80% in the study of outer membrane protein vaccines, but it cannot reach more than 90%.

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