Construction And Expression Of A Bivalent Membrane-anchored Expression DNA Vaccine Bearing Sj14FABP And Sj26GST Genes

Objective: To construct a eukaryotic membrane-anchored co-expression plasmid containing Sjc14FABP and Sjc26GST genes and identify their expression in vitro.Methods: Sj14 and Sj26 genes were obtained by RT-PCR with total RNA of Schistosoma japonicum adult worms as the template and cloned into eukaryotic expression plasmid pVAC to construct recombinant plasmid pVAC-Sj14 and pVAC-Sj26. Then two modified gene fragments were respectively amplified by PCR with templates of plasmid pVAC-Sj14 and pVAC-Sj26, which encode a nucleotide sequence of human interleukin-2 (IL-2)signal peptides in their 5'ends and that of membrane-anchored peptides of carboxyl-terminal of placental alkaline phosphatase (PLAP) in 3'ends. After that, the two modified genes were altogether cloned into a eukaryotic co-expression plasmid pIRES, resulting in another new recombinant plasmid pIRES-Sj26-Sj14. Then, their expression was identified through RT-PCR and indirect immunofluorescent assays (IFA) when the plasmid pIRES-Sj26-Sj14 was transfected into eukaryotic Hela cells.Results: Restriction enzyme analysis, PCR and sequencing results showed that the recombinant plasmid pVAC-Sj14, pVAC-Sj26, pIRES-Sj26-Sj14 were successfully constructed and the expression of modified Sj14 and Sj26 genes could be detected by RT-PCR and IFA.Conclusion: A bivalent membrane-anchored expression DNA vaccine bearing Sj14 and Sj26 genes was acquired, which layed foundations for later study of its immunogenicity and anti-affection efficiency.