Abstract:
Nucleotide sequences recognized and bound by DNA-binding proteins (DBPs) are critical to controlling and maintaining gene expression, replication, chromosome segregation, cell division, and nucleoid structure in bacterial cells. Therefore, determination of the binding sequences of DBPs is important not only to study DBP recognition mechanisms but also to understand the fundamentals of cell homeostasis. While ChIP-seq analysis appears to be an effective way to determine DBP binding sites on the genome, the resolution is sometimes not sufficient to identify the sites precisely. Here we introduce a simple and effective method named Genome Footprinting with high-throughput sequencing (GeF-seq) to determine binding sites of DBPs with single base-pair resolution. GeF-seq detects binding sites of DBPs as sharp peaks and thus makes it possible to identify the recognition sequence in each \"binding peak\" more easily and accurately compared to the common ChIP-seq.
摘要:
DNA结合蛋白(DBPs)识别和结合的核苷酸序列对于控制和维持基因表达至关重要。复制,染色体分离,细胞分裂,和细菌细胞中的类核结构。因此,确定DBPs的结合序列不仅对于研究DBP识别机制而且对于了解细胞稳态的基本原理很重要。虽然ChIP-seq分析似乎是确定基因组上DBP结合位点的有效方法,该决议有时不足以准确识别站点。在这里,我们介绍了一种简单有效的方法,称为高通量测序的基因组足迹(GeF-seq),以确定具有单碱基对分辨率的DBPs的结合位点。GeF-seq将DBP的结合位点检测为尖峰,因此与普通ChIP-seq相比,可以更轻松,更准确地识别每个“结合峰”中的识别序列。