稳定表达萤火虫荧光素酶的9L^(luc)细胞株的构建  

Constructing and applying of 9L^(luc) cells expressing firefly

作　　者：杨丽平[1] 赵敬湘[2] 李露斯[1] 袁红峰[2] 刘大庆[2] 王蕴芳[2] 南雪[2] 白慈贤[2] 裴雪涛[2] 

机构地区：[1]第三军医大学西南医院神经内科,重庆400038 [2]军事医学科学院输血研究所干细胞与再生医学研究室,北京100850

出　　处：《重庆医学》2007年第9期806-808,共3页Chongqing medicine

基　　金：国家重点基础研究发展规划(973)资助项目(2001CB509906和2005CB522702);国家"863"计划重大专项资助项目(2005AA219010)

摘　　要：目的构建稳定表达萤火虫荧光素酶的9Lluc细胞。方法通过酶切方法获得表达萤火虫荧光素酶的目的基因片段,将其连接到慢病毒表达质粒pBPLV,构建慢病毒表达载体pBPLV-Luc;将pBPLV-Luc转染到9L胶质瘤细胞,构建稳定表达萤火虫荧光素酶的9Lluc细胞并扩增培养。应用生物素荧光成像检测9Lluc细胞体内及体外表达。结果9Lluc细胞所释放的光子量均与细胞数量呈正相关。结论结果表明萤火虫荧光素酶已稳定转染入9Lluc细胞。Objective To construct 9L^luc cells expressing firefly luciferase. Methods The PGL3-basic plasmid was digested and the cDNA fragment encoding firefly luciferase was isolated; the gene of Luc was cloned into the lentivirus expression plasmid pB- PLV and construct pBPLV-Luc. The pBPLV-Luc were transfected into 9L, and obtained 9L^luc cells using flowcytometry.9L^luc cells were examined with bioluminescence imaging in vivo and in vitro. Results Bioluniescence was correlated to tumor cell number and luciferase was stable over time in vitro and in vivo. Conclusion The results indicated 9L^luc was constructed successfully.

关 键 词：隧病毒载体 荧光素酶 生物素荧光成像 

分 类 号：Q78[生物学—分子生物学]