Erkina Tamara Y, Erkine Alexandre M
College of Pharmacy and Health Sciences, Butler University, Indianapolis, IN, USA.
Methods Mol Biol. 2012;809:279-89. doi: 10.1007/978-1-61779-376-9_19.
Investigation of DNA-protein interactions is a key approach in understanding mechanisms of gene regulation. The method described allows detection of dynamic DNA-protein interactions occurring at gene promoters in living cells during the time scale of seconds and minutes. The combination of chromatin immunoprecipitation with real-time PCR allows for detection of changes in activator and co-activator content of any promoter during transcriptional activation. The described method is most applicable to investigation of processes resulting in nucleosome loss at gene promoters during the induction of transcription. The approach is also applicable to any dynamic process involving DNA-protein interactions.
DNA-蛋白质相互作用的研究是理解基因调控机制的关键方法。所描述的方法能够检测活细胞中基因启动子处发生的、时间尺度在秒和分钟范围内的动态DNA-蛋白质相互作用。染色质免疫沉淀与实时PCR相结合，能够检测转录激活过程中任何启动子的激活剂和共激活剂含量的变化。所描述的方法最适用于研究转录诱导过程中导致基因启动子处核小体丢失的过程。该方法也适用于任何涉及DNA-蛋白质相互作用的动态过程。
