Recombinant bacillus subtilis for displaying vibrio parahaemolyticus VP OmpK-MeL on spore surface, construction method and application

By constructing a recombinant Bacillus subtilis strain VPOmpK-MeL displaying Vibrio parahaemolyticus on its spore surface, the problem of the lack of highly effective oral vaccines in aquaculture has been solved. This approach achieves genetic stability and immune enhancement, simplifies the immunization procedure, and is suitable for the development of safe vaccines for small fish.

CN121674429APending Publication Date: 2026-03-17FUJIAN LUODONG BIOTECHNOLOGY CO LTD
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Patent Information

Application Number
CN202511945082.1
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2025-12-22
Publication Date
2026-03-17

AI Technical Summary

Technical Problem

There is a lack of highly effective and safe vaccines against Vibrio parahaemolyticus in current aquaculture, especially oral vaccines suitable for small fish. Furthermore, traditional vaccines have stability and safety issues, are difficult to administer, and have unstable protective effects.

Method used

A recombinant Bacillus subtilis spore surface displaying Vibrio parahaemolyticus VPOmpK-MeL was constructed. The OmpK antigen and MeL fusion protein were displayed on the surface of Bacillus subtilis spores using genetic engineering methods. Recombination was performed using the integrative plasmid pDG364 to ensure genetic stability. The spores were then transformed into B. subtilis 168 via homologous double cross transformation to achieve direct oral immunization.

Benefits of technology

This recombinant strain displays the VP OM protein on the spore surface, simplifying the immunization procedure, reducing costs, stimulating a specific immune response, enhancing the immune effect, protecting the intestinal barrier function, and reducing inflammatory response, making it suitable for commercial vaccine development.

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Abstract

The invention discloses a spore surface display vibrio parahaemolyticus VPOmpK-MeL recombinant bacillus subtilis as well as a construction method and application thereof. According to the recombinant bacillus subtilis, on the basis of an integrated plasmid pDG364, a recombinant vector which takes spore capsid protein CotY as anchoring protein and displays a fusion fragment of a VPOmpK antigen and bee venom peptide Melittin on the surface is constructed, recombinant protein is converted into a wild type bacillus subtilis 168 gene, and the obtained recombinant spore expressing OmpK-MeL protein has genetic stability; the bacterium directly displays antigen protein on the surface of a spore, does not need to be broken, and can be directly mixed with materials or drink water to immunize animals, so that the problems of difficult protein purification, tedious injection immunization process and the like are avoided; the spores have good stress resistance, so that the spores are easy to store and transport, and the cost is remarkably reduced; the strain can induce specific immune response, and the fusion expressed MeL plays an excellent role in enhancing immunity, so that a new way is provided for prevention and treatment of VP, and the strain can be applied to development of commercial vaccines.
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