Studies On Sjzfp1 Gene Of Schistosomisis Japunica

Schistosomisis japonica is a serious parasitic disease that causes serious healthy problem to both human and animals. It is reported that Microtus fortis is the only mammalian animals with native resistance to S. japonicum. Characterizing the target molecules of S. japonicum related to the schistosoma resistance of M. fortis might aid in identifying potential chemotherapeutic targets and/or vaccine candidates against schistosome infection.Sjzfp1, a zinc finger protein encoding gene, was obtained in precious work of the group of Pro. Liu from Shanghai Veterinary Research Institute, through screening a adult worm phage display cDNA library from worms of S. japonicum with fresh sera of Microtus fortis.Vaccine test of both phage display Sjzfp1 and recombinant fusion protein rSjzfp1/GST, showed that Sjzfp1 is an encouraging candidate antigen for development of an anti-schistosomiasis vaccine. However, It was very difficult to purify rSjzfp1/GST. In this paper, It was studied that cellular location, It was studied that protective immunity for expression of proteins of part genes of Sjzfp1and screening siRNA molecules, too.The 181bp-786 bp partial cDNA of Sjzfp1 was amplified by PCR, subcloned into pET-28a(+) vectors and express recombinant protein rSjzfp1-1/His in E.coli BL21(DE3) when induced with IPTG, the expression product was a 27.8 KDa fusion protein. Recombinant protein rSjzfp1-1/His was purified with Ni-NTA HisBind Resin.The purified rSjzfp1-1/His were used to vaccinate mice, it was obtained that reduced reductions of 17.85% for worm burden and 15.96% for hepatic egg burden.The full-length of Sjzfpl was amplified by PCR, subcloned into pEGFP-cl vectors, and it was transfected into 293T cells and expressing in 293T cells successfully. It was observed that pEGFP-c1-Sjzfp1 express in nucleus.The expression profiles of Sjzfp1 was determined by real-time PCR using the template cDNAs isolated from differential stages parasite. Real-time PCR analysis revealed that the Sjznf1 was expressed in all stages of the parasite, the highest expression was in cercariae and miracidia, and the highest expression in the stages in final host was in 42d-adult.In order to analyse expression profiles of Sjzfp1 in cercariae, RNA interference of this gene was primarily studied.three siRNAs were designed and used to interfere the expression of Sjzfp1 gene in cultured worms in vitro.The biological function of Szfp1 was deeply revealed in the study,that provid important basis the development of Sjzfp1 as candidate vaccine and drug targets.