重组慢病毒载体介导不同启动子驱动的绿色荧光蛋白在多种(不同)细胞中的表达影响  

Green florescence protein expression in various cells driven by different promoters among lentivirus

作　　者：张玲[1] 翁云层 张云 帅丽芳[3] 李婷婷[1] 王文敬[1] 李红卫[1] 赵卫[4] 黎诚耀[1] 

机构地区：[1]南方医科大学生物技术学院输血医学系,广东广州510515 [2]临朐县中医院骨外科,山东潍坊261000 [3]广州军区疾病预防控制中心,广东广州510515 [4]南方医科大学公共卫生与热带医学学院,广东广州510515

出　　处：《分子诊断与治疗杂志》2015年第3期161-165,共5页Journal of Molecular Diagnostics and Therapy

基　　金：国家自然科学基金(81301433)

摘　　要：目的比较不同启动子的慢病毒转导细胞后,在不同细胞系中驱动绿色荧光蛋白表达的效率高低。方法采用3种不同启动子的转移质粒,与包装质粒共转染293T细胞,转染60 h后,收集慢病毒上清。用等量的三种慢病毒液转导5种细胞系(293A、MOLT-4、PC3、DU145及RM1),72 h后,荧光显微镜下观察转导效果;流式细胞仪计数转导效率。结果不同启动子(Ubiquitin,EF1α,CMV)在5种细胞系中驱动绿色荧光蛋白的表达及转导效率不同。在293A和PC3细胞中,CMV为最强启动子,转导率分别为(94.83±2.87)%和(20.90±3.15)%;但在MOLT-4和DU145细胞中,EF1α为最强启动子,转导率分别为(74.27±2.14)%和(25.13±4.95)%;在RM1细胞中,Ubiquitin为最强启动子,转导率为(16.77±0.38)%。结论在慢病毒载体介导基因表达研究中,要考虑选取合理的细胞和启动子以获得高效的转导效率。Objective To compare the GFP expression efficiency of transduced cells driven by lend- virus with different promoters. Methods 293T cells were co-transfected with packaging plasmids and three kinds of transfer plasmids. Then the supernatant was collected after 60 hours. 5 cell lines （293A, MOLT-4, PC3, DU145 and RM1） were transduced by equal load of these lentivirus. 72 hours later, GFP expression was ob- served with florescence microscope and transduction efficency was counted by FASC. Results GFP expres- sion and transduction efficiency among 5 cell lines were different. Among 293A and PC3 cells, CMV promoter was the strongest one and transduction efficiency can achieve （94.83 ± 2,87）% and （20.90 ± 3.15）%, respec- tively. But among MOLT-4 and DU145 cells, EFloL promoter was the strongest one and transduction efficiency can achieve （74.27 ± 2.14）% and （25.13 ±4.95）%, respectively. In RM1 cells, Ubiquitin promoter was the strongest one and it can achieve （16.77 ±0.38）%. Conclusion In the study of gene expression mediated by lentivirus, proper cell lines and promoters should be selected to obtain high efficiency.

关 键 词：重组慢病毒 绿色荧光蛋白 

分 类 号：R450[医药卫生—治疗学]