日本血吸虫中国大陆株22.6kD膜相关蛋白基因的克隆与表达  

Cloning of Schistosoma japonicum Chinese Strain 22.6 kD Membrane-associated Protein(Sj-22.6)Gene and Its Overproduction in E.coli

作　　者：蔡学忠[1] 林矫娇 杨冠珍[2] 施福恢[1] 沈纬[1] 蔡幼民[1] 吴祥甫[2] 

机构地区：[1]中国农业科学院上海家畜寄生虫病研究所,上海20023 [2]中国科学院上海生物化学研究所

出　　处：《中国农业大学学报》1998年第S2期65-68,共4页Journal of China Agricultural University

基　　金：国家863生物高技术项目(102-07-04-03);总理专项基金(94-Y-40)部分经费资助

摘　　要：以日本血吸虫 mRNA 为模板,用 RT-PCR 法快速克隆到一大小约600bp 的 DNA 片段,DNA 序列分析证实,所扩增到的 DNA 片段中含有日本血吸虫22.6kD 膜相关蛋白(Sj-22.6(Ch))基因,将该基因重组到表达型质粒 pGEX-4T 中,表达的 GST 融合蛋白分子量约48kD,用谷胱甘肽琼脂塘凝胶亲和层析柱纯化的重组蛋白不仅纯度好,而且得率高,纯化产量可达40mg·L^(-1)培养物,免疫试验结果表明该重组蛋白具有良好的抗原性,为其在血吸虫病抗感染中的免疫作用研究创造了条件.A 576 bp DNA fragment was amplificated by RT-PCR,from Schistosoma japon- icum adult worm mRNA.Sequence analysis revealed that this fragment contained S.japon- icum Chinese strain membrane-associated protein(Sj-22.6)gene.Then this gene was cloned into the expression vector pGEX-4T,and subsequently expressed in Escherichia col- i.The recombinant GST-fusion protein was purified by glutathione agarose affinity chro- matography.Its molecular weight was about 48 kD.The yield of expression was around 40 mg·L^(-1)E.coli culture.The immunological test suggested that the recombinant protein had good antigenity,which could make a good basis for the research of its immunological function in Schistosomiasis.

关 键 词：22.6kD膜蛋白 日本血吸虫 基因克隆 基因表达 免疫作用 

分 类 号：S852.7[农业科学—基础兽医学]