Zhang Liang, Cheng Guo-Feng, Fu Zhi-Qiang, Jin Ya-Mei, Feng Xin-Gang, Yuan Chun-Xiu, Cai You-Min, Lin Jiao-Jiao
Shanghai Institute of Animal Parasitology, the Chinese Academy of Agricultural Science, Shanghai 200232, China.
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai). 2003 Dec;35(12):1099-104.
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.
基于东方田鼠自然抗血吸虫病现象，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，获得两个阳性克隆，进一步应用RACE技术扩增其中一个克隆，鉴定出一个具有开放阅读框（ORF）的cDNA片段。测序显示这是日本血吸虫的一个新基因，命名为SjMF4（日本血吸虫东方田鼠4）。然后分析了SjMF4的结构和功能基序。将该基因亚克隆到pET-28a(+)载体中；重组蛋白在Western印迹中显示出良好的抗原性。进一步将该基因亚克隆到真核表达载体pcDNA3中构建含SjMF4的DNA疫苗。小鼠免疫实验显示出显著的保护作用，重组质粒对日本血吸虫尾蚴攻击的减虫率为28.64%±3.82%，减卵率为21.73%±3.98%，均显著高于对照组。
