Grisolia V, Carlomagno M S, Bruni C B
J Bacteriol. 1982 Aug;151(2):692-700. doi: 10.1128/jb.151.2.692-700.1982.
The operator-distal genes hisBHAFI(E) of the Escherichia coli K-12 histidine operon were mapped on a DNA fragment 4,500 base pairs long. This fragment, originally present in a lambda transducing phage, was cloned in the vector plasmid pBR313. A restriction map was determined, allowing identification of the orientation of the genes in the fragment. The cloned genes were expressed in appropriate hosts, independent of the orientation of the DNA fragment, as shown by transformation tests and by enzyme assays of one of the gene products, hisB, histidinol phosphatase. An internal transcription initiation site was identified by isolation of the cellular RNA, hybridization to specific DNA probes, and mapping by S1 nuclease.
大肠杆菌K-12组氨酸操纵子的操纵基因-远端基因hisBHAFI(E)被定位在一段4500个碱基对长的DNA片段上。该片段最初存在于一个λ转导噬菌体中，被克隆到载体质粒pBR313中。确定了限制性酶切图谱，从而能够鉴定片段中基因的方向。如转化试验以及其中一个基因产物组氨醇磷酸酶（hisB）的酶活性测定所示，克隆的基因在合适的宿主中表达，且与DNA片段的方向无关。通过分离细胞RNA、与特异性DNA探针杂交以及用S1核酸酶进行定位，确定了一个内部转录起始位点。
