Skarstad K, Thöny B, Hwang D S, Kornberg A
Department of Biochemistry, Stanford University School of Medicine, California 94305-5307.
J Biol Chem. 1993 Mar 15;268(8):5365-70.
The replication origin of Escherichia coli (oriC) was probed for specific binding proteins, using the gel shift assay. A 33-kDa protein that binds specifically to the right border of oriC, next to the rightmost of the binding sites of the initiator protein, DnaA, was identified and purified. The stoichiometry of protein to DNA is about 6 to 1. Around 5000 monomers of the protein, named Rob (right oriC binding), are present per cell. The rob gene has been located near 99.8 min on the E. coli map, cloned, and overexpressed. The total protein sequence reveals strong homologies to several regulatory proteins with which it shares the helix-turn-helix motif.
利用凝胶迁移实验探究了大肠杆菌复制起点（oriC）的特异性结合蛋白。鉴定并纯化了一种33 kDa的蛋白，该蛋白特异性结合oriC的右边界，紧邻起始蛋白DnaA最右侧的结合位点。蛋白质与DNA的化学计量比约为6比1。每个细胞中大约存在5000个这种名为Rob（oriC右边界结合蛋白）的蛋白质单体。rob基因位于大肠杆菌染色体图谱上99.8分钟附近，已被克隆并过量表达。该蛋白质的完整序列显示出与几种具有螺旋-转角-螺旋基序的调控蛋白有很强的同源性。
