ParBF DNA binding profiles over various supercoiling level and on linear DNA

We report the genome-wide analysis from chromatin immunoprecipitated DNA (ChIP-sequencing) of the DNA binding pattern of ParBF (SopB) of plasmid F. This study, performed in E. coli and Salmonella typhimurium, investigates the impact of DNA supercoiling on ParBF DNA binding profiles in vivo. We found that variation in DNA supercoiling does not significantly affect the ParB DNA binding profiles even on linear DNA. Overall design: Chromatin immunoprecipitation DNA-sequencing for ParB of plasmid F (ParBF) in bacterial strains exhibiting various DNA supercoiling level, and on linearized plasmid F at several positions relative to the centromere parSF. Biological replicates are labelled IP1 and IP2. Anti ParBF antibody is affinity purified before used.

本研究报道了针对F质粒的ParBF（又称SopB）的DNA结合模式所开展的染色质免疫沉淀测序（ChIP-sequencing）全基因组分析。本研究在大肠杆菌（E. coli）与鼠伤寒沙门氏菌（Salmonella typhimurium）中开展，旨在探究DNA超螺旋对ParBF体内DNA结合谱的影响。研究结果表明，即便在线性DNA底物上，DNA超螺旋的变化也不会对ParBF的DNA结合谱产生显著影响。
总体实验设计：针对携带不同DNA超螺旋水平的细菌菌株中的F质粒的ParB蛋白（ParBF）开展染色质免疫沉淀测序，并针对相对于着丝粒序列parSF的多个位点的线性化F质粒进行该分析。生物学重复样本标记为IP1和IP2。实验所用的抗ParBF抗体在使用前经过了亲和纯化处理。