中文摘要：

目的构建WT1（Wilms＇tumor susceptibility gene,WT1）基因siRNA（small interference RNA,siRNA）载体,为进一步探讨WT1的生物学功能奠定基础.方法根据针对WT1 mRNA的有效的siRNA序列,设计合成两条shRNA（smallhair RNA,shRNA）的DNA模板单链,同时模板链两端分别设计不同的两个限制酶切位点.退火形成siRNA载体插入片断。用限制性内切酶将siRNA空载体线性化,T4连接酶将插入片断插入siRNA空质粒中.结果经酶切、PCR及测序鉴定确定构建成功.结论成功构建了WT1的干扰质粒,可对其生物学行为作进一步研究.

英文摘要：

Objective Establish a siRNA vector to go further research on the function of WT1 gene. Methods Two single strand DNA templates were designed according to the sequence of siRNA which has confirmed to be effective to knockdown WTI mRNA.When the DNA templates synthesized, two different restriction sites were superimposed ,respectively, to the two end of it.The insertion element formed after the DNA templates annealed. Make the blank plasmid linearization by use of restriction enzyme,then the insertion element was inserted into the blank plasmid by T4 ligase.Results Enzyme cutting,PCR and sequencing has confirmed that the construction is successful.Conclusion WTI siRNA plasmid was successfully constructed,and we could go further research on the function of WT 1 gene.