Recombine A Oral Live Vaccine Expression Rotavirus Antigen Gene VP4

Objective: To construct a recombinant plasmid of rotavirus antigen VP4(pBEX/VP4), detect the expression of VP4 gene, and test the safety of the expressed proten.Methods: (1)According to the sequence of murine rotavirus antigen gene VP4,34 primers were designed.The full-length VP4 gene was synthesized through overlapping PCR and cloned into a shuttle expression vector pBEX which originated from plasmid pGEX-5x-1.This vector was named pBEX/VP4(.2)Transformed the pBEX/VP4 into E coli DH5α.The product was tested by SDS-PAGE and its safety tested by ansa intestinalis test .Results: (1) Sequencing results shown that the production of PCR was correct ,and the recombinant plasmid was construced successfully. (2) VP4 gene could stable expressed in E coli DH5α,and the production of the VP4 has no toxicity of inducing intestinal juice secretory in SD rat by ansa intestinalis test.Conclusion: (1)Overlap PCR is a utility method to synthesize long-length gene without the template of the objective jene.(2) The expression of VP4 shown that the shuttle expression vector was constructed successfully ,and the production of VP4 which tested by ansa intestinalis test has no toxicity ,so ,it's the base of Transforming the pBEX/VP4 into the B. infantis and the studying of the new rotavirus oral vaccine .