Abstract：
Chromatin immunoprecipitation (ChIP) is used to analyze the targeting of a protein to a specific region of chromatin in vivo. Here, we present an instructive ChIP protocol for Arabidopsis imbibed seeds. The protocol covers all steps, from the sampling of imbibed seeds to the reverse crosslinking of immunoprecipitated protein-DNA complexes, and includes experimental tips and notes. The targeting of the protein to DNA is determined by quantitative PCR (qPCR) using reverse crosslinked DNA. The protocol can be further scaled up for ChIP-sequencing (ChIP-seq) analysis. As an example of the protocol, we include a ChIP-quantitative PCR (ChIP-qPCR) analysis demonstrating the targeting of PIF1 to the ABI5 promoter.
摘要：
染色质免疫沉淀（ChIP）用于分析体内蛋白质对染色质特定区域的靶向。这里,我们为拟南芥吸收的种子提出了一个有指导意义的ChIP协议。协议涵盖了所有步骤，从吸收种子的取样到免疫沉淀的蛋白质-DNA复合物的反向交联，并包括实验提示和笔记。通过使用反向交联的DNA的定量PCR(qPCR)测定蛋白质对DNA的靶向。该方案可以进一步扩大用于ChIP测序(ChIP-seq)分析。作为协议的一个例子，我们包括ChIP定量PCR（ChIP-qPCR）分析，证明PIF1靶向ABI5启动子。
