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日本血吸虫CAI基因的亚克隆表达与免疫保护效果研究

Study on the Subcloning, Expression and Immunoprotection with Schistosoma japonicum CAI Gene

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【作者】 罗秀菊袁仕善易新元曾宪芳张顺科唐连飞蔡春章洁Larry McReynolds

【Author】 LUO Xiu-ju~1,YUAN Shi-shan~1 , YI Xin-yuan~(1 *), ZENG Xian-fang~1 , ZHANG Shun-ke~1, TANG Lian-fei~1, CAI Chun~1, ZHANG Jie~1, Larry McReynolds~2 (1 Department of Parasitology, Xiangya School of Medicine, Central South University, Changsha 410078; 2 New England Biolabs, Inc. MA 01905 USA)

【机构】 中南大学湘雅医学院寄生虫教研室New England Biolabs 长沙410078长沙410078Inc.MA01905 USA

【摘要】 目的 克隆和表达日本血吸虫 (Schistosomajaponicum ,Sj)CAI基因 ,并对表达产物进行免疫保护效果测定 ,评价其作为血吸虫疫苗候选分子的潜能。 方法 将SjCAI基因亚克隆至pGEX 5X 3载体 ,转化入感受态大肠杆菌ER2 56 6 ,在异丙基 β D 硫代半乳糖苷 (IPTG)诱导下进行表达 ,用表达产物免疫小鼠 ,并设分别注射等体积的弗氏完全佐剂和PBS的两组对照组 ,观察免疫保护效果。 结果 在IPTG诱导下 ,表达载体中的SjGST基因与重组的SjCAI基因在大肠杆菌中获得了高效融合表达 ,将其免疫小鼠 ,诱导产生了 2 9.87%的减虫率和 6 3.71 %的减卵率。 结论 SjCAI基因亚克隆至pGEX 5X 3载体后可在大肠杆菌中高效表达 ,表达产物能诱导产生一定水平的抗Sj保护性免疫力

【Abstract】 Objective To subclone and express the new gene of Schistosoma japonicum (Sj) CAI and evaluate the immunoprotective effect of the recombinant molecule. Methods The cDNA of SjCAI gene was subcloned into expression vector pGEX-5X-3 to form recombinants which were then used to transform to E.coli strain ER 2566. Expression was induced by IPTG. The mice were vaccinated with the expressed protein and the immunoprotective effect was tested. Results Fusion protein of SjGST-CAI was highly expressed in E. coli as inclusion bodies. The worm reduction rate and the liver egg reduction rate in vaccination group of SjGST-CAI were 29.87% and 63.71%, respectively. Conclution SjCAI gene can be highly expressed in E .coli after subcloning into pGEX-5X-3 vector and the expressed fusion protein can induce immunoprotective effect against Sj in mice.

【基金】 湖南省科技厅资助项目 (oojzy 2 1 1 5 );WHO /TDR资助项目 (ID 980 2 68)~~
  • 【文献出处】 中国寄生虫学与寄生虫病杂志 ,Chinese Journal of Parasitology and Parasitic Diseases , 编辑部邮箱 ,2003年06期
  • 【分类号】R392
  • 【被引频次】2
  • 【下载频次】37
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