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抑制CDC25A表达的siRNA真核表达载体的构建

朱小东;李相德;曲颂;李龄;黎丹戎;张玮;李力;唐步坚;   

  1. 广西医科大学肿瘤医院;广西医科大学研究生学院;
  • 收稿日期:2009-06-25 修回日期:2009-06-25 出版日期:2009-06-25 发布日期:2009-06-25

Construction of eukaryotic expression vector of siRNA-inhibited CDC25A expression

ZHU Xiao-dong,LI Xiang-de,QU Song,et al.   

  1. (Affiliated Cancer Hospital of Guangxi Medical University,Nanning Guangxi 530021,China
  • Received:2009-06-25 Revised:2009-06-25 Online:2009-06-25 Published:2009-06-25

摘要: 目的在哺乳动物细胞中构建并鉴定抑制CDC25A表达的siRNA真核表达载体。方法根据CDC25A的基因序列,设计特异性的siRNA,将合成的siRNA核酸片段退火形成双链后连接到经Bam HI和HindⅢ双酶切后的psilencer4.1真核表达载体,命名为psilencer4.1-CDC25A以及psilencer4.1-Control,并进行酶切及测序鉴定。结果通过酶切鉴定和序列测定,成功地构建出抑制CDC25A表达的siRNA载体及其阴性对照载体。结论成功构建和验证了siRNA真核表达载体,为下一步研究奠定了良好的基础。

关键词: CDC25A, siRNA, 载体

Abstract: Objective To construct and identify an eukaryotic expression vector of siRNA-inhibited CDC25A expression in mammalian cells.Methods CDC25A-specific siRNA was designed and synthesized according to the sequence of CDC25A.After annealing the nucleic acid fragment of the siRNA to form the double-stranded,they were connected with the eukaryotic expression vector psilencer4.1 which was pre-digested with Bam HI and HindⅢ.Then the constructed vectors,named psilencer 4.1-CDC25A and psilencer4.1-Control,were digested and sequenced.Results The vector of siRNA-inhibited CDC25A expression and the negative control vector were constructed,sequenced and identified.Conclusion The successful construction and validation of the siRNA-inhibited CDC25A expression vector lays a good foundation for the next steps in CDC25A-related study.

Key words: CDC25A, siRNA, Vector