中文摘要：

目的：构建可受四环素诱导调控的高表达外源基因的真核表达体统A549pTevOn细胞系。用于F10基因的功能研究。方法：将pTet-On质粒用脂质体介导法转染A549细胞，利用G418的药物选择特性，对转染的A549细胞进行压力筛选后的克隆用有限稀释法进行单克隆化，单克隆分别扩增后，瞬时转染pTREquc（编码荧光素酶蛋白）质粒，Dox诱导表达后，检测荧光素酶活性，逐一筛选四环素调控高表达外源基因的A549pTet-On细胞株。结果：成功构建了一株受Dox调控的高表达低背景的A549pTet—On细胞株。结论：筛选的A549细胞为F10基因表达能被pTet-on诱导表达系统调控的细胞系，为研究F10基因功能提供了理想的细胞模型。适合在此基础上对F10基因进行深入的研究。

英文摘要：

Objective： To establish tetracycline controlled inducible system （pTet-on） in A549 cell. Methods： The A549 cells were transfected with pTet-on vector by using liposome transfection reagent. After 48 hours,the transfected cells were selected in medium containing G418, G418-resistant clones were isolated. All individual G418-resistant clones were screened by transient with pTRE-luc for clones with low background and high induction of luciferase in reponse to Dox. Results： One A549 cell line which exhibited levels of induction and high gene expression levels. Conclusion： The A549 cell cline can be used to highly express F10 gene and this pTet--on system is available for use in F10 gene function studies.