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Schistosomiasis is a worldwide infectious diseases, endangering the life and health of human beings, and impede social development. Schistosome DNA vaccine is a new approach to the prevention of schistosome disease, Schistosoma japonicum DNA vaccines have made great progress. This article cloned Schistosoma japonicum antigen gene; gene fusion antigen gene; build Schistosoma japonicum DNA vaccines; Schistosoma japonicum DNA vaccines fermentation process, separation and purification processes; Schistosoma japonicum DNA vaccines in vivo, in vitro expression of feasibility . Achieved the following: 1) antigen gene cloned four kinds of Schistosoma japonicum sj23, sjFABP, sjGAPDH, sj26 verify positive clones after restriction enzyme digestion, sequencing; 2) using fusion PCR experimental technology, constructed two fusion genes sj23.FABP and sj26.GAPDH; 3) subcloned four kinds the the the Japanese schistosome antigen gene sj23, sjFABP, sjGAPDH sj26 and two kinds of Schistosoma japonicum antigen gene fusion gene sj23.FABP of sj26.GAPDH inserted into the eukaryotic expression vector pVIVO2 polyclonal sites, building a 14 Schistosoma japonicum DNA vaccines; 4) to optimize the fermentation conditions of Schistosoma japonicum DNA vaccines: the use of single-factor experiments, optimal carbon source for glycerol carbon sources, nitrogen sources, the optimum nitrogen source is soybean juice; Best K2HPO4 add 20%; screened three important influence factor: soybean juice, beef juice, glycerin \\soybean juice of 4.21%, an initial pH of 7.0; 5) the establishment of the preparation process of Schistosoma japonicum DNA vaccines: gel filtration to remove RNA and contaminating proteins by affinity and ion chromatography to remove the open-loop plasmid DNA affinity adsorption of endotoxin. The final product after quality assessment, agarose gel electrophoresis is not visible RNA bands the hybrid protein mass fraction of 0.11%, supercoiled plasmid DNA content of 93.4%, the total heat source the content for 0.85EU / (mg by microinjecting the plasmid containing). The indicators have reached the technical requirements of the State Food and Drug Administration for medical grade DNA vaccine. 6) through the indirect fluorescent immunoassay, 14 Schistosoma japonicum DNA vaccine were all able to at the antigen level in the muscle of mice, and MCF-7 eukaryotic cells successfully expressed.
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