Expression of group A rotavirus VP7 gene in recombined Lactobacillus

ObjectiveTo constmct vector pEDM 27/5-P7 by RT-PCR and transfoun it into Lacotobacillus for the construction of genetic engineering vaccine against rotavirus.MethodsThe VP7 gene fragment was isolated and recoveried by RT-PCR, and tmnsfoun edinto pEDM 27/5.The gene was transformed into Lactobacillus through extraction from the plasmid.After lactose induction, the vp7 was detected with SDS-PACE and Western-blot at 4, 8 and 12 hours.ResultsThe VP7 gene was transformed into pEDM 27/5 successfully, and the susceptibity bacteria was recombined by pEDM 27/5-vp7.After lactose induction at 4, 8, 12 hours, the protein (28KD)was detected with SDS-PACE and Western-blot.The expressed protein was 2.30%,5.12%,5.38% of total protein of the bacteria, respectively by scanning, and the expression increased with the increment of the time.The protein could combined it specially combined with anti-vp7 protein.ConclusionThe recombinant Lactobacillus expresses immunocompetent vp7 protein constantly, and could be used for further development of genetic engineering vaccine against rotavirus.