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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

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【作者】 赵志荣刘淼雷黎朱绍春汪学龙沈际佳

【Author】 ZHAO Zhi-rong,LIU Miao,LEI li,ZHU Shao-chun,WANG Xue-long,SHEN Ji-jia(Department of Parasitology,Anhui Medical University,Hefei 230032,China)

【机构】 安徽医科大学病原生物教研室安徽医科大学病原生物教研室 合肥230032现工作单位安徽省马鞍山市疾病预防控制中心合肥230032

【摘要】 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。

【Abstract】 To construct the expression vector of RHA interference(RNAi) system for gene coding for the Mago nashi-like 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.

【关键词】 日本血吸虫Magonashi样蛋白RNAisiRNAshRNA
【Key words】 Schistosoma japonicumMago nashi like proteinRNAisiRNAshRNA
【基金】 安徽省教育厅自然科学基金重点项目(2005KJ045ZD)
  • 【文献出处】 中国人兽共患病学报 ,Chinese Journal of Zoonoses , 编辑部邮箱 ,2006年08期
  • 【分类号】R383.2
  • 【被引频次】4
  • 【下载频次】113
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