Sela S, Clark-Curtiss J E
Department of Biology, Washington University, St. Louis, MO 63130.
Gene. 1991 Feb 1;98(1):123-7. doi: 10.1016/0378-1119(91)90114-q.
The putative promoter region of the 16S ribosomal RNA-encoding gene (rRNA) of Mycobacterium leprae was cloned and characterized in Escherichia coli. A 932-bp HaeIII restriction fragment, containing the 5' end of the 16S rRNA gene and flanking upstream region, was cloned in front of a promoterless reporter gene in the shuttle vector, pMH109, to generate the plasmid, pYA1101. This clone exhibits promoter activity both in Gram-(E. coli) and Gram+ (Bacillus subtilis) bacteria. Sequence analysis and primer extension experiments with mRNA derived from the M. leprae clone were used to determine the structure and the location of the promoter, as well as the transcription start point in E. coli. The promoter region contains sequences that resemble the -35 and -10 consensus sequences found in many bacteria. A region located 34 bp distal to the promoter is a putative rRNA processing signal, based on sequence homology with processing signals involved in the maturation of the rRNA precursor in B. subtilis and several Mycoplasma species.
麻风分枝杆菌16S核糖体RNA编码基因（rRNA）的假定启动子区域在大肠杆菌中被克隆并进行了特性分析。一个932 bp的HaeIII限制性片段，包含16S rRNA基因的5'端及侧翼上游区域，被克隆到穿梭载体pMH109中无启动子的报告基因前，构建成质粒pYA1101。该克隆在革兰氏阴性菌（大肠杆菌）和革兰氏阳性菌（枯草芽孢杆菌）中均表现出启动子活性。通过对来自麻风分枝杆菌克隆的mRNA进行序列分析和引物延伸实验，确定了启动子的结构、位置以及在大肠杆菌中的转录起始点。启动子区域包含与许多细菌中发现的-35和-10共有序列相似的序列。基于与枯草芽孢杆菌和几种支原体属中rRNA前体成熟所涉及的加工信号的序列同源性，启动子下游34 bp处的一个区域是一个假定的rRNA加工信号。
