大鼠NgR特异性siRNA慢病毒载体的构建与鉴定

目的将siRNA技术应用于NgR（Nogo受体），构建出有效的NgR特异性siRNA慢病毒载体。方法按照E1bashir等设计原则和siRNA的要求，设计、合成4对siRNA，并将其与双酶切慢病毒载体pGCSIL-GFP连接，转化，挑取阳性克隆进行PCR鉴定，阳性克隆送测序。结果4对NgR特异性siRNA与双酶切慢病毒载体pGCSIL-GFP连接成功。结论4对NgR特异性siRNA慢病毒载体的构建成功，为进一步合成病毒载体及观察NgR特异性RNA干扰抑制大鼠NgR mRNA表达对脑白质损伤的修复作用奠定了基础。

Construction and Identification of Specific siRNA Rat NgR Gene Lentivirus Vector

Objective Construction of effective NgR (Nogo receptor) specific siRNA lentivirus vector by the application of siRNA technique. Method Four pairs of siRNAs were designed and synthesized based on the Elbashir principle and siRNA intrinsic character and the ligation products of the siRNAs with double digested Lentivirus vector pGCSIL-GFP were transformed. Colonies were PCR screened and positive clones with putative in-frame ligation were further sequenced. Result The ligation of four pairs of NgR specific siRNAs to the double digested Lentivirus vector pGCSIL-GFP was successful. Conclusion The successful construction of four NgR specific siRNA lentivirus vectors shed light on other virus vector constructions as well as further study of the repair function of the NgR specific RNA interference on rat brain white matter damage by inhibiting NgR mRNA expression.