DMRIE-C、Lipofectamine2000和TransMessenger转染HCV RNA效率比较  

A comparison of DMRIE-C,Lipofectamine2000 and TransMessenger in their efficiency of transfection with HCV RNA

作　　者：高博[1] 贾帅争[1] 彭剑淳[1] 王怡[1] 詹林盛[1] 许金波[1] 

机构地区：[1]军事医学科学院野战输血研究所,北京100850

出　　处：《军事医学》2011年第7期517-520,共4页Military Medical Sciences

基　　金：国家自然科学基金资助项目(30801414);北京市自然科学基金资助项目(7082071)

摘　　要：目的比较和优化DMRIE-C、Lipofectamine2000和TransMessenger3种转染试剂的转染效率,选择适合建立丙型肝炎病毒（HCV）复制子或感染克隆的转染试剂和条件。方法将含有萤火虫荧光素酶报告基因的HCVRNA,分别通过上述3种转染试剂转染Huh7和Huh7.5.1细胞系,于转染24h后裂解细胞,测定荧光素酶活性。2~3周后,结晶紫染色细胞集落。结果在Huh7和Huh7.5.1细胞中,DMRIE-C比Lipofectamine2000和TransMes-senger的转染效率高。在Huh7细胞中,4μlDMRIE-C转染试剂组的荧光素酶活性比其他组高（P〈0.05）。在Huh7.5.1细胞中,3μlDMRIE-C转染试剂组的荧光素酶活性比其他组高（P〈0.05）。在细胞集落形成实验中,DMRIE-C组的细胞克隆数多于其他两种转染试剂组。结论在Huh7和Huh7.5.1细胞中,DMRIE-C比Lipo-fectamine2000和TransMessenger的转染效率高。当建立HCV复制子或感染克隆试验时,既要选择合适的RNA转染试剂,又要考虑细胞生长状态的影响。Objective To compare and optimize the transfection efficiency of DMRIE-C,Lipofectamine2000 with HCV RNA and TransMessenger with HCV RNA and then to choose proper transfection reagents for establishment of HCV replicon or infectious cell models.Methods The HCV RNA containing the firefly luciferase gene were transfected into Huh7 cells and Huh7.5.1 cells using DMRIE-C,Lipofectamine2000 and TransMessenger,respectively.At 24 h post-transfection,the transfected cells were lysed and the luciferase activity was assayed.At 2-3 weeks post-transfection,visible colonies were stained by crystal violet.Results The transfection efficiency of DMRIE-C was better than that of Lipofectamine2000 and TransMessenger in both Huh7 cells and Huh7.5.1 cells.In Huh7 cells,the firefly luciferase activity of 4 μl DMRIE-C transfection group was higher than in other groups（P0.05）.In Huh7.5.1 cells,the firefly luciferase activity of 3 μl DMRIE-C transfection group was higher than in other groups（P0.05）.In colony forming assay,the colony number of DMRIE-C group was larger than that of Lipofectamine2000 group and TransMessenger group in Huh7 and Huh7.5.1 cells.Conclusion The transfection efficiency of DMRIE-C is better than that of Lipofectamine2000 and TransMessenger in both Huh7 and Huh7.5.1 cells.Proper transfection reagents and growth status of cells need to be considered when HCV replicon or infectious cell models are established.

关 键 词：丙型肝炎病毒(HCV) RNA 荧光素酶类 萤火虫 转染 

分 类 号：R373.21[医药卫生—病原生物学]