中文摘要：

目的利用Tet-On诱导可调控系统构建表达淀粉样前体蛋白突变基因（APPswe）细胞模型。方法构建重组质粒p TRE-APPswe-Flag,利用慢病毒将其包装转染SH-SY5Y细胞。Puromycin筛选72 h后,通过强力霉素诱导,Western blot检测验证,免疫荧光检测APPswe表达情况。结果强力霉素诱导后,筛选的细胞Western blot检测Flag标签蛋白表达明显,免疫荧光显示APPswe蛋白表达增加;撤掉强力霉素后,APPswe蛋白表达减少。结论成功构建了可调控Tet-OnAPPswe表达SH-SY5Y细胞系统模型。

英文摘要：

Objective To establish an inducible APPswe expression cell line using Tet-On system. Methods The recombinant plasmid p TRE-APPswe-Flag was packaged by lentivirus and then transinfected SH-SY5 Y cells. After puromycin selection for 72 h, SH-SY5 Y cells were induced by doxcycline（Dox）. Expression of Flag tag and APPswe was determined by Western blot and immunofluorescent analysis, respectively. Results Expression of Flag and APPswe in SH-SY5 Y cells was upregulated after Dox induction, while expression of APPswe was downregulated after Dox removal. Conclusion An inducible expression of APPswewas successful established in SH-SY5 Y cells using Tet-on system.