Hall C V, Yanofsky C
J Bacteriol. 1981 Dec;148(3):941-9. doi: 10.1128/jb.148.3.941-949.1981.
From a Clark-Carbon plasmid containing trpS, the structural gene for the tryptophanyl-transfer ribonucleic acid synthetase of Escherichia coli, we subcloned a 2.6-kilobase fragment that has trpS and its neighboring regions. The location and orientation of trpS in the deoxyribonucleic acid insert was determined by deoxyribonucleic acid sequencing. In vitro transcription experiments and S1 nuclease mapping studies with in vivo message established that transcription is initiated at the same site in vivo and in vitro, approximately 58 base pairs upstream from the trpS coding region. We also describe the construction of an inphase trpS-lacZ gene fusion that is under the control of the trpS promoter and encodes a hybrid protein with beta-galactosidase activity.
从一个含有大肠杆菌色氨酰 - 转移核糖核酸合成酶结构基因trpS的克拉克 - 卡本质粒中，我们亚克隆了一个包含trpS及其相邻区域的2.6千碱基片段。通过脱氧核糖核酸测序确定了trpS在脱氧核糖核酸插入片段中的位置和方向。体外转录实验以及利用体内信使进行的S1核酸酶图谱分析研究表明，体内和体外转录均在同一位置起始，该位置在trpS编码区上游约58个碱基对处。我们还描述了一个同相位trpS - lacZ基因融合体的构建，该融合体受trpS启动子控制，并编码具有β - 半乳糖苷酶活性的杂合蛋白。
