C-erbB-2特异性siRNA逆转录病毒载体的构建与鉴定

Construction and Identification of Recombinant Retroviral Vector of siRNA Specific for C-erbB-2

目的构建并鉴定C-erbB-2特异性siRNA逆转录病毒载体。方法体外合成含针对C-erbB-2特异基因序列的寡核苷酸,并定向克隆入逆转录病毒载体RNAi-ReadypSIREN-RetroQ。结果构建的重组逆转录病毒载体可以被限制性内切酶MIuI、BamHI、EcoRI切开,电泳可显示相应条带,经测序其序列与设计的一致。结论成功构建了C-erbB-2特异性siRNA逆转录病毒载体pRetroQ/C-erbB-2/siRNA-1、pRetroQ/C-erbB-2/siRNA-2、pSIREN-RetroQ/C-erbB-2/siR-NA-3,为进一步研究p185基因沉默奠定了基础。

Objective To construct and identify recombinant retroviral vectors of siRNA specific for C-erbB-2. Methods We synthesized oligonucleotides for C-erbB-2 in vitro and cloned them into retroviral vector RNAi-Ready pSIREN-RetroQ. Results The recombinant retroviral vectors digested by MIu I,BamH I,EcoR I showed the corresponding bands, and their sequences were the same to the anticipation. Conclusion The recombinant retroviral vectors of pSIREN-RetroQ/C-erbB-2/siRNA-I,pSIREN-RetroQ/C-erbB-2/siRNA-2 and pSIREN-RetroQ/C-erbB-2/siRNA-3 specific for C-erbB-2 were constructed successfully,which contributed to the study of p185 gene silencing.