Regulatory Roles and Networks of the Whole Set of Transcription Factors in Escherichia coli

基因组的表达是通过与转录因子的相互作用来控制基因组内有限数量的RNA聚合酶的分布。本研究的目的是确定所有300个转录因子在模型原核生物大肠杆菌中的调节作用。1)开发了体外基因组SELEX筛选系统，确定了300个转录因子的调控靶基因。2)我们还开发了体内nip芯片系统，并确定了转录因子在大肠杆菌基因组内的细胞内分布。3)通过检测它们在转录因子存在和不存在情况下的表达来检测由此预测的调控靶点。4)我们通过定量免疫印迹分析确定了所有300个转录因子的细胞内水平。5)综合所有结果，我提出：(1)所有300个转录因子的调控靶点清单；(2) 300个转录因子的相互作用网络，共同形成复杂的层次调控网络。

The genome expression is determined by controlling the distribution of a limited number of RNA polymerase within the genome through interaction with transcription factors. The aim of this research was to identify the regulatory role of all 300 transcription factors in the model prokaryote, Escherichia coli.1) We developed the in vitro Genomic SELEX screening system and determined the regulation target genes for all 300 transcription factors.2) We also developed the in vivo NIP-chip system and determined the intracellular distribution of transcription factors within the E. coli genome.3) The regulation targets thus predicted were examined by measuring their expression in the presence and absence of transcription factors.4) We determined the intracellular levels of all 300 transcription factors by using quantitative immune-blot analysis.5) Taking all the results together, I have proposed :(1) the list of regulation targets for all 300 transcription factors ; and(2) the interaction network of 300 transcription factors, altogether forming complex hierarchic regulation networks.