为了寻找日本血吸虫(Schistosomajaponicum,Sj) 新的疫苗候选基因并进行免疫效果研究,用Sj雌虫抗原免疫家兔制备血清,对Sj成虫cDNA文库进行免疫筛选,将获得的新基因(命名为Sj-F1,GenBank登录号为AY261995) 克隆入原核表达载体pTWIN1和真核表达载体pcDNA3,经PCR、限制性酶切筛选和鉴定阳性重组子. 将pTWIN1/Sj-F1质粒转化大肠杆菌ER2566,在低温和低IPTG浓度下诱导表达可溶性重组融合蛋白(rSj-F1/intein2),并经SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白质印迹(Western blot)分析鉴定. 将pcDNA3/Sj-F1质粒转化大肠杆菌ER2502,大量制备DNA疫苗. 用重组融合蛋白和DNA疫苗免疫小鼠,末次免疫后2周用Sj尾蚴进行攻击感染. 感染后42天剖杀冲虫,计算减虫率和减卵率. 感染前采血用ELISA法检测抗体. 免疫保护效果测定显示:重组蛋白疫苗以FCA作佐剂经皮下免疫和以壳聚糖作佐剂经粘膜免疫分别获得了28.07%、24.69%的减虫率和48.30%、46.38%的减卵率;DNA疫苗(pcDNA3/Sj-F1)单独免疫获得了18.47%的减虫率和35.06%的减卵率;用DNA疫苗启动免疫后用重组蛋白疫苗经皮下加强免疫,减虫率和减卵率分别提高到了40.42%和56.17%;用DNA疫苗启动免疫后用重组蛋白疫苗经黏膜加强免疫,减虫率和减卵率增高更明显,分别提高到了42.38%和62.8 In order to search for new vaccine candidate genes of Schistosoma japonicum (Sj) and to study the immune effect, Sj female antigens were used to immunize rabbits to prepare serum. The Sj adult cDNA library was screened by immunization. The new gene (named as Sj- F1, GenBank accession number AY261995) was cloned into the prokaryotic expression vector pTWIN1 and the eukaryotic expression vector pcDNA3, and the recombinant plasmid was screened and identified by restriction enzyme digestion with restriction endonucleases. The plasmid pTWIN1 / Sj-F1 was transformed into Escherichia coli ER2566, Expression of soluble recombinant fusion protein (rSj-F1 / intein2) was induced at low IPTG concentration and identified by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis.The pcDNA3 / Sj-F1 plasmid E.coli ER2502 was transformed into E.coli to prepare a large amount of DNA vaccine.The mice were immunized with the recombinant fusion protein and DNA vaccine and challenged with SjCercariae 2 weeks after the last immunization.The insects were killed 42 days after infection to calculate the worm reduction and egg reduction rates Antibodies were detected by ELISA before infection.Immunoprotection results showed that 28.07% and 24.6% of recombinant protein vaccine were immunized with FCA as adjuvant and mucosal immunization with chitosan as adjuvant respectively 9% worm reduction rate and 48.30% reduction rate of 46.38%; DNA vaccine (pcDNA3 / Sj-F1) alone achieved 18.47% reduction in worm reduction rate and 35.06% The recombinant protein vaccine was boosted subcutaneously, and the worm reduction rate and the egg reduction rate increased to 40.42% and 56.17%, respectively. After immunization with recombinant DNA vaccine and DNA vaccine, , Respectively, increased to 42.38% and 62.8