【摘要】 目的:应用携带有报告基因-βGal的质粒对于使用L ipofectam ine 2000转染大鼠肝脏细胞及中国仓鼠肾脏成纤维细胞条件进行优化。方法:在96孔板上按不同比例混合L ipofectam ine 2000和质粒DNA后转染大鼠肝脏细胞及中国仓鼠肾脏成纤维细胞,成功转染的细胞因可表达β-Gal可分解底物X-gal在染液的存在下可使细胞呈蓝色,故计算蓝色细胞的比例即可了解转染的效率。应用方差分析的方法进行统计分析。结果:对于BHK细胞,当DNA与L ipofectam ine 2000的比例为0.27∶0.8(μg∶μL)时转染效率最高,为(43.2±9.3)%;对于BRL细胞,DNA与L ipofectam ine 2000的比例为0.10∶0.3(μg∶μL)时转染效率最高,可达(5.7±1.3)%。结论:L ipofectam ine 2000可有效转染哺乳动物细胞,不同细胞有不同的最佳转染条件。更多还原

【Abstract】 Objective:To optimize the Lipofectamine 2000 transfection conditions for rat BHK cells and Chinese hamster BRL cells using β-Gal vectors.Methods: Lipofectamine 2000 were mixed in varied ratios with plasmid DNA to transfect the rat BHK cells and Chinese hamster BRL cells on a 96-well plate.Successfully transfected cells were stained blue due to expression of β-Gal degradable substrates(X-gal),enabling estimation of the transfection efficacy by counting the blue cells under microscope.ANOVA was performed for statistical test.Results: The maximal efficacy was(43.2±9.3)% in BHK cells when β-Gal vector and Lipofectamine 2000 were mixed in a ratio of 0.27∶0.8(μg∶μL),and was(5.7±1.3)% in BRL cells when β-Gal vector and Lipofectamine 2000 were mixed in a ratio of 0.10∶0.3(μg∶μL).Conclusion: Lipofectamine 2000 appears to be useful for the transfection of mammal cells;however,the transfection conditions should be optimized for individual cell types.更多还原