Cloning,Expression And Biological Function Analysis Of Genes IM Of Schistosoma Japonicum

Schistosomiasis is a serious threat and widespread zoonotic disease.In rencent years, schistosomiasis control has made remarkableachievements in China. However medication alone can not control theprevalence of schistosomiasis, to enhance screening and vaccinedevelopment, schistosomiasis vaccine candidate molecules isundoubtedly the schistosome disease prevention and control ofimportant initiatives. In mammals, IM (inositol monophosphate) to theinsulin lipid second messenger, the PI (3,4,5) P3hydrolyzed to PI (3,4) P2.However, in the IM gene function in the parasite rarely reported. thedevelopment and maturation of Schistosoma mechanisms, screeningof new anti-schistosome vaccine candidate molecules are verysignificance. Screening out of IM, to carry out the following studies.In this study, first cloning gene SjIM of schistosomiasis japonicum,GenBank accession number is FN319326.1,by blasting obtained thefull-length cDNA sequence of SjIM.Through bioinformatics analysis, TheOpen Reading Frame for SjIM were8344bp, encoding277amino acid,theoretical molecular weight of30.6Daltons. Real-time PCR analysisfrom the worms of various stages of S. japonicum revealed that themRNA level of SjIM was highest in the35days schistosomula.Successfully constructed pET28a (+)-SjIM recombinant prokaryoticexpression plasmid, recombinant protein in E.coli (DE3) in solubleexpression, molecular weight of30.6kD, Western blotting analysis showed that the fusion protein with good antigenicity. Purifiedrecombinant protein SjIM/HIS immunized BALB/c mice, compared withthe control group, recombinant proteins were induced in mice SjIM aworm reduction rate of48.76%and41.29%of the liver egg reductionrate, a significant difference (p <0.05). The change of the specific IgGand subtypes IgG1, IgG2a was measured by ELISA; the induced specificIgG and subtypes IgG1, IgG2a was maintained in a high level. It is showthat SjIM had the potential for the anti-schistosomal drugs.Successfully constructed the recombinant eukaryotic expressionplasmid pVAX1-SjIM, recombinant pVAX1-SjIM in BALB/c mice induceda28.94%worm reduction rate of39.07%of the liver and egg reductionrate, significant difference (p <0.05), after recombinant pVAX1-SjIMimmunizing mice three times, high titers of specific IgG antibodies wasinduced..The change of the specific IgG was measured by ELISA.preliminary results show that, compared with the control group.. All the results will assist us to understand more functions about SjIMand provide the basis for screening new Schistosomiasis vaccinecandidate molecules and drug targets.