日本血吸虫thioredoxin基因的克隆和表达  

The Cloning and Expression of Thioredoxin of S.japonicum

作　　者：邵筱[1] 余新炳[1] 吴忠道[1] 王海[1] 梁柏年[1] 李宝华[1] 

机构地区：[1]中山大学基础医学院寄生虫学教研室,广州510080

出　　处：《热带医学杂志》2004年第2期126-129,共4页Journal of Tropical Medicine

基　　金：国家自然科学基金(No.30060783);国家教育部博士点基金(No.200045)。

摘　　要：目的结合分子生物学和生物信息学方法筛选鉴定日本血吸虫新基因。方法从日本血吸虫(Schistosomajaponicum,大陆株)成虫cDNA文库中获取表达序列标签(expressedsequencetag,EST),用电子拼接的方法延伸序列,用NCBI提供的BLASTx程序和Genbank数据库进行同源性分析以筛选基因;设计特异性引物从日本血吸虫成虫mRNA中扩增筛选基因并预测和分析;扩增产物克隆到原核表达载体并表达。结果筛选出日本血吸虫Thioredoxin全长基因并对其进行了序列分析,克隆全长cDNA至PET原核载体并表达成功。结论结合EST、电子延伸和传统的分子生物学方法是高效筛选S.japonicum功能基因的有效策略。Objective To screen and identify novel genes from Schistosoma japonicum. Methods Expressed sequence tag(EST) from the cDNA library of S.japonicum was elongated by in silicon elongation system. The elongated ESTs were analyzed by BLASTx to find similar sequence in Genbank. According to the information of elongated sequence, design special primers to amplify the full length sequence of interested gene, and then, predict the function, clone and express the gene. Result The full length gene encoding Sj thioredoxin was cloned into PET vector and the recombinant protein whose molecular was about 12000u was expressed as well as identified by SDS PAGE. Conclusion It prove a effective method for obtaining important functional genes to combine the approach of molecular biology and bioinformatics.

关 键 词：日本血吸虫 THIOREDOXIN 基因克隆 基因表达 硫氧还蛋白 

分 类 号：R383.24[医药卫生—医学寄生虫学] R394[医药卫生—基础医学]