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四环素及其衍生物诱导表达的pTet-On肺癌细胞系A549的建立 预览 被引量:1

Establishment of A pTet-on Pulmonary Carcinoma Cell Line Regulated by Tetracycline and Doxycycline
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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。 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.
作者 曹晓敏 林仲秋 庞战军 全松 邢福祺 Cao Xiaomin, et al (Department of Obstetrics and Gynecology, Second Affiliated Hospital of Zhongshan University, Zhongshan 510120)
出处 《数理医药学杂志》 2009年第2期 158-160,共3页 Journal of Mathematical Medicine
基金 基金项目:国家自然科学基金资助项目(编号30470658,30672234)
关键词 四环素 F10基因 基因调控 tetracycline and derivative regulated expression F10 gene
作者简介 现工作单位:广东省中山市人民医院 曹晓敏(1973-),女,在站博士后,研究方向:生殖内分泌。 通讯作者:邢福祺
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