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

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）％。%Objective To compare the GFP expression efficiency of transduced cells driven by lenti-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, EF1α 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.