In vivo organization of bacterial transcription initiation complexes at a genome-scale [TSS-seq]

RNA polymerase (RNAP) is essential for the transcription of genetic information encoded in genomes in all three domains of life. To determine the precise organization of bacterial transcription initiation complexes (TIC) in vivo, we exploited high-throughput sequencing to the DNA obtained from exonuclease-treated immunoprecipitates of the TIC. It reveals that sigma (s) factor is mostly engaged in RNAP holoenzyme up to 13-14 nucleotides from transcription start site during abortive initiation. Overall design: TSS-seq (+TEX), and TSS-seq (-TEX) were generated in duplicates by using Illumina MiSeq

RNA聚合酶（RNA polymerase, RNAP）是生命三大域中基因组编码遗传信息转录过程的必需因子。为精准解析体内细菌转录起始复合物（transcription initiation complexes, TIC）的精细组织形式，我们对经外切酶处理的TIC免疫沉淀物中获取的DNA开展了高通量测序。分析结果显示，在流产式起始阶段，σ因子（sigma factor）大多结合于距转录起始位点13至14核苷酸的RNA聚合酶全酶上。实验设计概述：采用Illumina MiSeq平台对TSS-seq (+TEX)与TSS-seq (-TEX)进行双重复建库与测序。