日本血吸虫新基因sjf4的克隆、表达及功能分析

Liang Zhang, Guo-Feng Cheng, Zhi-Qiang Fu, Ya-Mei Jin, Xin-Gang Feng, Chun-Xiu Yuan, You-Min Cai, Jiao-Jiao Lin
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引用次数: 0

摘要

基于东方田鼠天然抗血吸虫病的现象,利用正常东方田鼠血清免疫筛选日本血吸虫(中国株)成虫cDNA文库,获得2个阳性克隆,应用RACE技术对其中1个克隆进行扩增,鉴定出具有ORF的cDNA片段。测序结果显示该基因为日本血吸虫新基因,并将其命名为sjf4 (Schistosoma japonicum Microtus fortis 4),分析了sjf4的结构和功能基序。将该基因亚克隆到pET-28a(+)载体中;Western blot结果显示重组蛋白具有良好的抗原性。将该基因亚克隆到真核表达载体pcDNA3中,构建含sjf4的DNA疫苗。小鼠免疫实验表明,与对照相比,重组质粒对日本血吸虫尾蚴攻击的虫减少率为28.64%+/-3.82%,卵减少率为21.73%+/-3.98%。
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[Cloning, expressing and functional analysis of SjMF4, a novel Schistosoma japonicum gene].

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.

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