中文摘要：

目的：构建小鼠成红细胞白血病病毒E26癌基因同系物1（V-Ets avian erythroblastosis virus E26 oncogene homolog1,Ets-1）重组腺病毒,并验证其对小鼠原代胰岛及胰岛β细胞系INS-1的感染活性和Ets-1蛋白表达活性。方法：以小鼠胰岛β细胞系MIN6的cDNA为模板,PCR扩增Ets-1基因的编码区序列（coding sequence,CDS）区,纯化后经限制性内切酶切割和连接反应插入到pAdTrack-CMV穿梭质粒上,构建成pAdTrack-CMV-Ets-1质粒。将该质粒与腺病毒骨架质粒pAdEasy-1在大肠杆菌BJ5183中进行同源重组,鉴定出阳性克隆。扩增并抽提阳性克隆质粒,用PacⅠ酶切线性化后转染293A细胞,经包装得到AdV-Ets-1重组腺病毒。相应对照病毒AdV-GFP由相同方法构建。腺病毒经纯化后,感染小鼠原代胰岛及INS-1细胞,并用Western blot检测Ets-1蛋白表达水平。结果：腺病毒构建成功,并具有高感染率及高蛋白表达能力。结论：成功构建了小鼠Ets-1腺病毒,为进一步研究Ets-1基因在原代胰岛中的功能提供基础。

英文摘要：

Objective：To construct a recombinant adenovirus that express mouse Ets-1 in primary cultured mouse islets. Methods：To generate pAdTrack-CMV-Ets-1 shuttle vector,the CDS region of Ets-1 was amplified by PCR,and then purified and cloned into the pAdTrack-CMV. pAdTrack-CMV-Ets-1 was recombined with back-bone pAdEasy-1 in BJ5183 bacteria. The resulting vector was transfected into QBI-293 A cells to generate recombinant adenovirus. After been amplified and purified,the recombinant adenovirus were used to infect primary cultured mouse islets and INS-1 cells. The protein levels of Ets-1 were determined by Western blotting assay. Results：The Adv-Ets-1 was established successfully and proved to be high infective and possess a high expression potential.Conclusion：The Ets-1 recombinant adenovirus was successfully constructed,which provided a foundation for further study of the function of Ets-1 in primary islets.