Expression and immunogenicity analysis of accessory colonization factor A from Vibrio alginolyticus strain HY9901

Abstract

The accessory colonization factor A (ACFA) of Vibrio alginolyticus plays an important role in the efficient colonization of the bacterium and is potential candidates for vaccine development. In present study, the acfA gene was cloned, expressed and purified. Western blot analysis revealed protein recognition with the native ACFA in different V. alginolyticus strains. To analyze the immunogenicity of the recombinant ACFA, Lutjanus erythropterus Bloch were immunized by intraperitoneal injection, and the results demonstrated that the recombinant ACFA produced an observable antibody response in all sera of the vaccinated fish. The differential expressions of RAG1 gene in various tissues of L. erythropterus were analyzed by fluorescent quantitative real-time PCR, and the results showed the RAG1 mRNA expression was significantly up-regulated in thymus, head kidney and spleen tissue. Furthermore, the protective property of recombinant ACFA was evaluated through challenge with six heterogeneous virulent V. alginolyticus strains, and the immunohistochemical analysis in different tissues after challenge with V. alginolyticus. The results showed L. erythropterus vaccinated with recombinant ACFA were more tolerant of the infection by virulent V. alginolyticus strains. The data indicate that the recombinant ACFA could provide heterologous protection for the different virulent V. alginolyticus strains.