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《Acta Microbiologica Sinica》 2008-11
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Expression and Immunogenicity Analysis of Recombinant Plasmid pW425et-Vp7 of Porcine Rotavirus A in Lactobacillus

Jingtao Hu, Chunfeng Wang (College of Animal Science, Jilin Agricultural University, Changchun 130118, China)  
Objective We tested the antigenicity of porcine rotavirus Vp7 gene expressed in Lactobacillus. MethodsWe 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.
【Fund】： 国家“863计划”(2003AA24112002、2006AA10A205 2007AA10Z322);; 国家自然科学基金(0200199、30671573);; 吉林省科技厅项目(20020220)~~
【CateGory Index】： S852.5
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