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目的:应用RNA干扰(RNAi)技术调节大鼠c-jun基因在Cos-7细胞中的表达。方法:分别构建大鼠c-jun基因的RNA干扰载体和真核表达GFP(绿色荧光蛋白)载体,将两者共转染Cos-7细胞,镜下观察大鼠C-Jun-GFP融合蛋白的表达,应用Western blot方法检测抑制效率。结果:酶切和测序结果表明,大鼠c-jun基因的RNA干扰载体和真核表达GFP载体构建成功,镜下及Western blot共转染结果均显示随着RNA干扰载体浓度的增加C-Jun-GFP融合蛋白表达量逐渐减少。结论:在Cos-7细胞中应用RNAi技术成功调节大鼠c-jun基因的表达。
OBJECTIVE: To use RNA interference (RNAi) technology to regulate the expression of c-jun gene in Cos-7 cells. Methods: RNA interference vector and GFP (green fluorescent protein) vector of rat c-jun gene were constructed respectively and co-transfected into Cos-7 cells. The expression of C-Jun-GFP fusion protein Expression, Western blot detection of inhibition efficiency. Results: The results of enzyme digestion and sequencing showed that the RNA interference vector and the eukaryotic GFP vector of rat c-jun gene were successfully constructed. The co-transfection results of microscopy and Western blot showed that with the increase of RNA interference vector concentration, C-Jun -GFP fusion protein expression decreased gradually. Conclusion: The expression of c-jun gene in rat Cos-7 cells was successfully regulated by RNAi technology.