血吸虫新抗原基因SjMF4的克隆、表达及功能分析  

Cloning, Expressing and Functional Analysis of SjMF4, a Novel Schistosoma japonicum Gene

作　　者：张亮[1] 程国锋[1] 付志强[1] 金亚美[1] 冯新港[1] 苑纯秀[1] 蔡幼民[1] 林矫矫[1] 

机构地区：[1]中国农业科学院上海家畜寄生虫病研究所、农业部动物寄生虫学重点实验室，上海200232

出　　处：《生物化学与生物物理学报》2003年第12期1099-1104,共6页

基　　金：上海市科技发展基金资助项目 (No .2 0 0 14 90 90 0 2 )~~

摘　　要：根据东方田鼠天然抗日本血吸虫病的现象 ,首次利用东方田鼠健康血清结合羊抗小鼠IgG3抗体免疫筛选日本血吸虫 (中国大陆株 )成虫cDNA表达型文库 ,获得两个阳性克隆 ,采用RACE技术对其中一cDNA片段进行扩增 ,获一含ORF的基因片段。序列分析表明该基因为一日本血吸虫新基因 ,命名为SjMF4 (SchistosomajaponicumMicrotusfortis 4 ,SjMF4 )。利用ExPASy的ScanProsite软件对此基因编码的蛋白质结构和功能域进行了分析。把该基因克隆到原核表达载体pET 2 8a(+) ,Western印迹显示表达产物具有良好的抗原性。又构建了真核表达质粒pcD NA3 SjMF4重组DNA疫苗 ,小鼠实验表明可诱导一定的保护作用。Based on the phenomenon of the natural anti-schistosomiasis in Microtus fortis, the sera from normal Microtus fortis were employed to immunoscreen the cDNA library of adult Schistosoma japonicum (Chinese strain), two positive clones were obtained, RACE technique was further applied to amplify one of the clones, and a cDNA fragment with an ORF was identified. Sequencing revealed that it was a novel gene of Schistosoma japonicum, and it was named SjMF4 (Schistosoma japonicum Microtus fortis 4). Then the structure and functional motifs of SjMF4 were analysed. The gene was subcloned into pET-28a(+) vector; the recombinant protein showed good antigenicity in Western blotting. The gene was further subcloned into eukaryotic expression vector pcDNA3 to construct the DNA vaccine containing SjMF4. Immune experiments in mice showed significant protection that the recombinant plasmid did induce 28.64%±3.82% worm reduction and 21.73%±3.98% egg reduction than controls against the Schistosoma japonicum cercaria challenge.

关 键 词：血吸虫 抗原基因 SjMF4 克隆 表达 东方田鼠 

分 类 号：S852.7[农业科学—基础兽医学]