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以荧光素酶基因为目的基因,应用Lipofectamine介导的瞬时转染方法,将pTet-on调控质粒和pTRE2hyg-luciferase表达质粒共转染小鼠胚胎干细胞,滴加系列浓度的DOX后,观察其诱导效应.结果显示,随DOX浓度升高,荧光素酶的活性也逐渐增强;而未转染细胞与转染细胞未加诱导剂时,其RLU值均接近空白对照.表明转染Tet-on基因表达系统的胚胎干细胞对DOX诱导有较好的反应性,目的基因的表达与DOX具有严格的剂量依赖关系.
Using luciferase gene as a target gene, pTet-on regulatory plasmid and pTRE2hyg-luciferase expression plasmid were cotransfected into mouse embryonic stem cells using Lipofectamine-mediated transient transfection. After a series of concentrations of DOX were added dropwise, the induction was induced The results showed that the luciferase activity increased gradually with the increase of DOX concentration, while the RLU value of untransfected cells and transfected cells without adjuvant was close to the blank control, indicating that transfection with Tet-on gene The expression system of embryonic stem cells had good reactivity to DOX induction, and the expression of the target gene was in a dose-dependent manner with DOX.