IFI16基因反义干扰表达载体的构建与鉴定

摘要

为了构建可在人喉癌细胞中稳定表达 IFI16基因短发夹RNA（shRNA）的表达载体，设计合成的IFI16基因shRNA片段，连接到经BamH I和 EcoR I双酶切的pGreenPuroTM shRNA表达载体中，连接产物转化大肠杆菌后挑取几个抗性菌落，用PCR技术初步进行鉴定，经PCR初步鉴定为重组质粒的一个重组子DNA用测序进一步鉴定，测序结果显示成功构建了 IFI16基因shRNA的表达载体pGreenPuro-IFI16 shRNA 。%To construct the vector that can stably express IFI16 shRNA in human laryngocarcinoma cells ,the well designed and correctly synthesized IFI16 shRNA fragment was used to ligate into the pGreenPuro TM shRNA expressing vector that predigested with BamH I and EcoR I .Subsequently ,the recombinant vector was used to transform the E .coli and then the PCR was performed to identify the resistant colonies .Besides ,the identified recombinant DNA was subjected to sequence for the further identification .Finally ,the IFI16 shRNA expressing vector named pGreenPuro-IFI16 shRNA was correctly constructed as circumstantiated by the sequencing result .