Nogo受体基因干扰RNA表达载体的构建

Construction of iRNA expression vector of Nogo receptor gene

目的构建Nogo受体基因干扰RNA表达载体,为促进中枢神经轴突的再生、探索临床治疗脊髓损伤新途径奠定基础。方法GeneBank中NgR的序列,应用www.invitrogen.com网站的设计软件设计、合成NgR miRNA的相应Oligo DNA,与BLOCK-iT Poll miR RNAi Expression Vector相连接,转化感受态Eco.li TOP10。结果限制性内切酶BamHⅠ和XhoⅠ酶切、测序鉴定重组载体BLOCK-iT Poll miR RNAi/NgR,显示NgR miRNA相应的Oligo正确克隆入BLOCK-iTPoll miR RNAi Expression Vector。结论成功构建Nogo受体基因干扰RNA表达载体。

Objective To construct iRNA expression vector of Nogo receptor gene. Accordingly it can establish foundation for promoting reclamation of central nerve axis and exploring new path of the treatment of spinal cord injury in clinic. Method The sequence of NgR was found in Gene Bank. We designed and synthesized Oligo DNA corresponding with NgR miRNA using the software in www. invitrogen, corn.The Oligo DNA was connected with BLOCK-iT Poll miR RNAi ＇Expression Vector, And was tramfomled into conlpetence Eco. li TOP10. Result the custom-crafted Vector, BLOCK-iT Poll miR RNAi / NgR, was Sequenced and identified using BanaH Ⅰand Xho Ⅰ . It is discovered that the Oligo DNA of NgR miRNA had been cloned into BLOCK-iT Poll miR RNAi Expression Vector correctly. Conclusion iRNA expression vector of Nogo receptor gene is constructed successfully.