Kim Soonok, Mitchell Thomas K
Department of Plant Pathology, The Ohio State University, Columbus, OH, USA.
Methods Mol Biol. 2011;722:121-31. doi: 10.1007/978-1-61779-040-9_8.
To attempt to gain an understanding of the molecular underpinnings of disease, many researchers have turned to expression profiling of genes during various stages of host recognition, entry, invasive growth, and host responses. While these studies have proven valuable, a deeper level of knowledge of the control circuitry affecting observed gene expression profiles can lead to a better understanding of the host pathogen interaction. Transcription factors are key switches in signal transduction circuits regulating gene expression. One powerful method to define target sequence specificity for this important group of transcription regulators is chromatin immunoprecipitation (ChIP) with microarray chips (chip), commonly called ChIP-chip. A more recent variation of this technique is ChIP-seq where DNA sequencing replaces the microarray chip. Here, we describe how we elucidated the binding sites for the Magnaporthe oryzae Ca(2+)/calcineurin-dependent transcription factor MoCRZ1 with the ChIP-chip approach.
为了试图了解疾病的分子基础，许多研究人员已转向在宿主识别、进入、侵袭性生长和宿主反应的各个阶段对基因进行表达谱分析。虽然这些研究已证明具有价值，但对影响观察到的基因表达谱的控制电路有更深入的了解可以更好地理解宿主与病原体的相互作用。转录因子是调节基因表达的信号转导电路中的关键开关。定义这一重要转录调节因子组的靶序列特异性的一种强大方法是使用微阵列芯片（芯片）进行染色质免疫沉淀（ChIP），通常称为ChIP-chip。该技术的一个更新变体是ChIP-seq，其中DNA测序取代了微阵列芯片。在这里，我们描述了我们如何使用ChIP-chip方法阐明稻瘟病菌Ca(2+)/钙调神经磷酸酶依赖性转录因子MoCRZ1的结合位点。
