日本血吸虫含EF手性结构域分子序列的分析和初步鉴定

SEQUENCE ANALYSIS AND PRELIMINARY IDENTIFICATION OF MOLECULES WITH EF-HAND DOMAIN OF SCHISTOSOMA JAPONICUM

为分离鉴定日本血吸虫含EF手性结构域分子及分析其序列结构特征,首先找到含有EF-hand结构域的分子,按照序列一级结构进行分类,之后进行生物信息学分析。然后用PCR方法以虫卵和成虫cDNA文库为模板扩增分类后的分子,构建重组质粒,诱导蛋白表达并纯化,选取14个纯化蛋白用Western blot进行初步免疫原性鉴定。结果269个原始含EF-hand结构域分子按照序列一级结构分为70个;成功表达和纯化了49个含EF手性分子,进化树分析将其分为9类;Western blot显示,14个纯化蛋白均可被日本血吸虫感染小鼠阳性血清识别。本研究结果为下一步采用这类分子进行动物保护性效果评价以及保护性免疫机制的研究奠定了基础。

To isolate and identify the molecules with EF-hand domain of Schistosoma japonicum and analyse the sequence structural characteristics of them, the molecules with EF-hand domain were firstly found and classified according to the primary structure, and then analyzed by bioinformatics. PCR was used to amplify the classified molecules with the S.japonicum adult and egg cDNA library. The recombinant plasmids were constructed and the protein with EF-hand domain were expressed and purified. Western blot was used to identify the preliminary immunogenicity of 14 purified proteins. Two hundred and sixty nine original molecules with EF-hand domain were divided into 70 molecules. Forty nine molecules with EF-hand domain were expressed and divided into 9 categories by phylogenetic tree. Western blot analysis showed that 14 purified proteins were specifically recognized by the positive serum from mice infected with S.japonicum. The study provided the basis for further study of immune protection of the molecules.