Construction and expression of the recombinant plasmid pVIVO2-IL-12-Sj-14FABP for Schistosoma japonicum

To construct the recombinant plasmid co expressing 14 kDa fatty acid binding protein from Schistosoma japonicum(Sj14FABP) and IL 12 and to express this plasmid in mice, the total RNA was extracted from adult worms of S.japonicum;the gene encoding Sj14FABP was amplified by RT PCR and digested with restriction endonucleases BamHI and EcoRI for a directional gene cloning to the vector pVIVO2 IL 12. The resulting construct was identified by PCR, restriction endonuclease digestion and sequence analysis. This recombinant plasmid was then used to immunize BALB/c mice and the local muscle tissue was analyzed by means of indirect fluorescence assay (IFA). It was found that a 440 bp fragment was amplified through RT PCR, and the constructed recombinant plasmid containing the sequence of Sj14FABP gene was obtained after PCR, enzyme digestion and sequence analysis. It was proved by IFA that the recombinant plasmid pVIVO2 IL 12 Sj14FABP was expressed in muscle cells.It concludes that the recombinant plasmid pVIVO2 IL 12 Sj14FABP has been successfully constructed and expressed in mice.