Grisolia V, Riccio A, Bruni C B
J Bacteriol. 1983 Sep;155(3):1288-96. doi: 10.1128/jb.155.3.1288-1296.1983.
The entire histidine operon of Escherichia coli K-12 was cloned in the vector plasmid pBR313, and a complete restriction map of the operon was determined. By using subclones, complementation tests, and enzyme assays, we were able to make a correlation between the physical map and the genetic map of the operon. We determined the sequence of a fragment of DNA 665 base pairs long, comprising the distal portion of the hisC gene, the proximal portion of the hisB gene, and the internal transcription initiation site hisBp. The efficiency of this promoter was assessed under different physiological conditions by cloning the DNA fragment in a recombinant vector system used to study transcriptional regulatory signals. The precise point at which transcription initiates was determined by S1 nuclease mapping.
将大肠杆菌K-12的整个组氨酸操纵子克隆到载体质粒pBR313中,并确定了该操纵子的完整限制性图谱。通过使用亚克隆、互补试验和酶分析,我们能够建立该操纵子物理图谱与遗传图谱之间的关联。我们测定了一段665个碱基对长的DNA片段的序列,该片段包含hisC基因的远端部分、hisB基因的近端部分以及内部转录起始位点hisBp。通过将该DNA片段克隆到用于研究转录调控信号的重组载体系统中,在不同生理条件下评估了该启动子的效率。通过S1核酸酶图谱分析确定了转录起始的精确位点。