南方医科大学学报Issue(12)：1761-1765,5.DOI:10.3969/j.issn.1673-4254.2013.12.11
细胞角蛋白8 GFP/Puro双标干涉慢病毒系统的构建及对细胞凋亡的影响
Construction of a GFP/Puro double-labeled lentiviral vector containing CK8 interfering RNA and its effect on cell apoptosis in vitro
摘要
Abstract
Objective To construct a GFP/Puro double-labeled lentiviral expression vector for CK8 silencing and assess the effects of CK8 silencing on cell apoptosis. Methods The siRNA sequences of CK8 were inserted into the lentiviral expression vector GV248 and transfected into 293T cells with the packaging plasmids PMD and SPA. The lentivirus was collected at 24 and 36 h post-transfection. Flow cytometry was used to detect the virus titer and the positive cells were selected with puromycin. The knockdown of CK8 was examined by Western blotting. The effect of CK8 down-regulation on cell apoptosis induced by cisplatin was detected with Annexin V/PI staining. Results and Conclusion We successfully constructed CK8 interference lentiviral vector and obtained a stable cell line with CK8 knock-down that was sensitive to cisplatin-induced apoptosis.关键词
细胞角蛋白8/GFP/Puro双标慢病毒/RNAi/凋亡/顺铂Key words
CK8/GFP/Puro double-marker lentiviral vector/RNA interference/apoptosis/cisplatin引用本文复制引用
金延超,尹荣华,郑巍薇,孔祥祯,詹轶群,杨晓明,李长燕..细胞角蛋白8 GFP/Puro双标干涉慢病毒系统的构建及对细胞凋亡的影响[J].南方医科大学学报,2013,(12):1761-1765,5.基金项目
国家自然科学基金（31170712）Supported by National Natural Science Foundation of China (31170712) （31170712）
