日本血吸虫tetraspanins胞外环2编码基因的克隆表达及其免疫原性的初步研究  

Researches on Cloning and Expression of the Gene Encoding Schistosoma japonicum Tetraspanins Extracellular Loop 2 and Its Immunogenicity

作　　者：付译节[1,2] 杨莉[1] 刘康[1] 袁创[1] 陈县城[1] 魏于全[1] 

机构地区：[1]四川大学华西医院生物治疗国家重点实验室,成都610041 [2]成都大学医护学院

出　　处：《四川大学学报（医学版）》2008年第6期890-894,共5页Journal of Sichuan University(Medical Sciences)

摘　　要：目的克隆和表达日本血吸虫(中国大陆株)tetraspanins胞外环2编码基因(Sj-tsp-2),初步研究其免疫原性。方法通过全基因合成方式获得目的基因片段,构建重组质粒pET32a-Sj-tsp-2,转化大肠杆菌BL21(DE3)感受态细胞,经IPTG诱导表达,在非变性条件下纯化融合蛋白。结果通过核苷酸测序证实重组表达质粒构建成功。SDS-PAGE结果表明,融合蛋白和目的肽段与预计大小基本一致。Western Blotting和ELISA结果说明,该融合蛋白具有良好的免疫原性。免疫定位分析显示,Sj-TSP-2主要在日本血吸虫体被表达。结论本实验成功克隆和表达了日本血吸虫Sj-tsp-2基因,并纯化获得其融合蛋白,为进行动物保护性实验奠定了基础。Objective To clone and express the gene （Sj-tsp-2） encoding extracellular loop 2 （EC-2） of tetraspanins （TSPs） of Schistosoma japonicurn （Chinese strain ）, and then to study the antigenicity and immunogenicity of this protein. Methods Synthesized Sj-tsp-2 gene was cloned into prokaryotic expression vector pET32a to generate pET32a-Sj-tsp-2 recombinant plasmid, and transformed into competent E. coli BL21 （DE3）. After inducing with IPTG, the expressed fusion protein was purified under nondenaturing conditions. Results The recombinant was confirmed by sequence analysis. The size of fusion protein and interest peptide was accords with the theoretical value by SDS-PAGE analysis. Additionally, the results of Western Blotting and ELISA demonstrated that the expressed protein had good immunogenicity. More importantly, we confirmed that TSP-2 is mainly located on the surface of S. japonicurn. Conclusion The successful expression and purification of recombinant protein Trx-Sj-TSP-2 will be very helpful for the further study of its protection role in animals.

关 键 词：日本血吸虫 TETRASPANINS 克隆 融合蛋白 免疫原性 

分 类 号：R383.24[医药卫生—医学寄生虫学] R392.11[医药卫生—基础医学]