Postle K, Good R F
Proc Natl Acad Sci U S A. 1983 Sep;80(17):5235-9. doi: 10.1073/pnas.80.17.5235.
The nucleotide sequence of a cloned section of the Escherichia coli chromosome containing the tonB gene has been determined. Transcription initiation and termination sites for tonB RNA have been determined by S1 nuclease mapping. The tonB promoter and terminator resemble other E. coli promoters and terminators; the sequence of the tonB terminator region suggests that it may function bidirectionally. The DNA sequence specifies an open translation reading frame between the 5' and 3' RNA termini whose location is consistent with the position of previously isolated tonB::IS1 mutations. The DNA sequence predicts a proline-rich protein with a calculated size of 26.1-26.6 kilodaltons (239-244 amino acids), depending on which of three potential initiation codons is utilized. The predicted NH2 terminus of tonB protein resembles the proteolytically cleaved signal sequences of E. coli periplasmic and outer membrane proteins; the overall hydrophilic character of the protein sequence suggests that the bulk of the tonB protein is not embedded within the inner or outer membrane. A significant discrepancy exists between the calculated size of tonB protein and the apparent size of 36 kilodaltons determined by sodium dodecyl sulfate/polyacrylamide gel electrophoresis.
已测定了大肠杆菌染色体中包含tonB基因的一个克隆片段的核苷酸序列。通过S1核酸酶图谱分析确定了tonB RNA的转录起始和终止位点。tonB启动子和终止子与其他大肠杆菌启动子和终止子相似;tonB终止子区域的序列表明它可能双向发挥作用。DNA序列在5'和3' RNA末端之间指定了一个开放的翻译阅读框,其位置与先前分离的tonB::IS1突变的位置一致。根据所使用的三个潜在起始密码子中的哪一个,DNA序列预测出一种富含脯氨酸的蛋白质,计算大小为26.1 - 26.6千道尔顿(239 - 244个氨基酸)。tonB蛋白预测的NH2末端类似于大肠杆菌周质和外膜蛋白的经蛋白酶切割的信号序列;蛋白质序列的整体亲水性表明tonB蛋白的大部分不嵌入内膜或外膜中。tonB蛋白的计算大小与通过十二烷基硫酸钠/聚丙烯酰胺凝胶电泳测定的36千道尔顿的表观大小之间存在显著差异。