Regulation of the senescence responsive genes by nucleosomal change

在本研究中，我们验证了5-溴脱氧尿嘧啶是否可以改变人类HeLa细胞中对5-溴脱氧尿嘧啶快速反应的PAI-1基因启动子区域的核小体结构。我们开发了“引物延伸法”来精确分析核小体在DNA上的定位。用微球菌核酸酶对染色质DNA进行酶切，然后用特定引物进行连接和PCR，发现了被认为是由核酸酶酶切产生的特定条带。通过努力消除非特异性条带，我们可以精确地检测核小体结构的变化。

In this study, we verified whether 5-bromodeoxyuridine can change nucleosomal structure in the promoter region of the PAI-1 gene quickly responsive to 5-bromodeoxyuridine in human HeLa cells. We developed "the primer extension method" to precisely analyze the positioning of nucleosomes on DNA. Digestion of the chromatin DNA with micrococcal nuclease followed by linker ligation and PCR with specific primers revealed specific bands that are thought to be derived from digestion with the nuclease. With efforts to eliminate non-specific bands, we came to a situation where we can precisely detect a change in the nucleosommal structure.