【摘要】 目的检测红藻氨酸(KA)处理后的大鼠大脑皮质神经元中p53蛋白与p21基因启动子的结合情况。方法原代培养大鼠大脑皮质神经元经KA或对照溶剂(双蒸水)处理后,采用染色质免疫沉淀(ChIP)及聚合酶链反应(PCR)技术检测神经元中p53蛋白与p21基因启动子区p53反应元件1(RE1)及反应元件2(RE2)的结合情况,Western印迹检测免疫沉淀中的p53蛋白以验证ChIP抗体特异性。结果 KA处理组,p53抗体沉淀的染色质中包含p21基因启动子区含有p53 RE1的DNA序列,免疫沉淀样品中含有p53蛋白。结论 KA处理后神经元内p53蛋白与p21基因启动子区p53 RE1结合增加。更多还原

【Abstract】 【Objective】To detect the binding of p53 to p21 gene promoter in rat cerebral cortical neurons exposed to kainate(KA).【Methods】Primary cultured rat cerebral cortical neurons were exposed to KA or control solvent(ddH2O) respectively,then chromatin immunoprecipitation(ChIP) and PCR analysis were used to detect the binding of p53 to p53 response element 1(RE1) or p53 response element 2(RE2) of p21 gene promoter in neurons.To testify the specificity of p53 antibody in ChIP,Western blotting was used to detect p53 in immunoprecipitated samples.【Results】In KA group,the p53 RE1-containing DNA fragments in p21 gene promoter were found in p53 antibody immunoprecipitated chromatins,and p53 was detected in immunoprecipitated samples.【Conclusions】The binding of p53 to p53 RE1 of p21 gene promoter increases in neurons exposed to KA.更多还原