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Abstract
目的 建立一种染色质免疫沉淀技术(ChIP)分析肝细胞基因表达的转录调控方法.方法以肝脏特异性基因磷酸烯醇式丙酮酸激酶(PEPCK)为待测基因,以IgG和次黄嘌呤鸟嘌呤磷酸核糖基转移酶(HPRT)及髓磷脂基础蛋白(MBP)作为内参基因,采用ChIP验证不同代谢状态下cAMP应答元件结合蛋白(CREB)对PEPCK基因启动子区的结合情况.结果在抗CREB抗体免疫沉淀的DNA模板中含有一明显增强的DNA片段,即禁食状态下PEPCK基因启动子区域CREB的结合增加4倍,证明DNA片段中含有与PEPCK基因启动子区结合的CREB序列.结论研究证实,CREB蛋白在体内可结合于PEPCK基因的启动子区域,并参与该基因表达的转录调控。
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