Expression and Immunogenicity Analysis of Recombinant Plasmid pW425et-Vp7 of Porcine Rotavirus A in Lactobacillus

Abstract:

Abstract: [Objective] We tested the antigenicity of porcine rotavirus Vp7 gene expressed in Lactobacillus. [Methods] We studied the condition of electroporation and transformed the plasmid pW425et-Vp7 into thyA gene-mutant L. acidlophilus. Then the positive clone was selected and its expression was detected by SDS-PAGE and Western blot. In our test, the BALB/c mice were drenched by the above-mentioned recombinant strain and SIgA was detected by ELISA. In addition, we also tested the protection of the recombinant strain through attacking porcine rotavirus to the mice immunized (content of virus was 15 mg/mL). [Result] Vp7 gene expressed in Lactobacillus with approximate 40.77 kDa fusion protein was observed by SDS-PAGE. The protein had the reactionogenicity with the antibody of porcine rotavirus through Western blot. We also detected the specific SIgA from the mice immunized by ELISA. Moreover, we observed that only 23% mice had diarrhea after attacking porcine rotavirus, which was immunized by recombinant strain. [Conclusion] Vp7 gene was expressed in Lactobacillus successfully. It had good immunogenicity and protection against porcine rotavirus. Moreover, the test further proved that the recombinant strain could be applied in the study on porcine rotavirus vaccine in the future.