日本血吸虫Mago nashi基因RNA干扰系统的构建及鉴定

Construction and identification of the RNA interference system for Schistosoma japanicum Mago nashi gene

目的构建针对日本血吸虫Mago nashi样蛋白基因的RNA干扰的表达载体。方法设计及化学合成日本血吸虫Mago nashi样蛋白基因的短发夹结构的寡核苷酸,通过退火成双链DNA片段,将其与经限制性内切酶BglⅡ和HindⅢ双酶切的pSUPER质粒连接,构建表达shRNA的重组质粒。结果经酶切及测序证实针对日本血吸虫Mago nashi样蛋白基因的RNA干扰的表达载体pSUPER构建成功。结论成功构建针对日本血吸虫Mago nashi样蛋白基因的RNA干扰的表达载体,为进一步研究对日本血吸虫Mago nashi样蛋白基因RNA干扰作用及该基因功能奠定基础。

To construct the expression vector of RHA interference （RNAi） system for gene coding for the Mago nashilike protein of Schistosoma japonicum, the oligonucleotides targeting to Mango nashi-like protein of Schistosoma japonicum were devised and chemically synthesized. They were annealed to form double-strand DNA and plasmid pSUPER was digested by Bgl Ⅱ and Hind Ⅲ to form linear vector. Finally, the annealed DNA segment was inserted into the linear vector to construct the recombinant plasmid of RMAi, The recombinant plasmid was then identified with restriction enzymes and comfirmed by sequencing analysis. It is established that the RNA interference system for Mago nashi-like protein of S. japonicum is successfully constructed.