Débarbouillé M, Raibaud O
J Bacteriol. 1983 Mar;153(3):1221-7. doi: 10.1128/jb.153.3.1221-1227.1983.
The malPQ operon, one of the three operons of the maltose regulon, is positively controlled by the product of gene malT. The starting point for malPQ transcription was deduced from experiments which involved a hybridization of in vivo-synthesized malPQ mRNA with adequate DNA probes, followed either by a digestion of nonhybridized DNA (S1 nuclease mapping) or by an extension of the hybridized probe (reverse transcriptase mapping). In the wild-type strain, this starting point was 37 nucleotides upstream from the initiation codon for malP. This analysis was also performed on a double mutant which contained both a 13-base pair deletion and a 3-base pair insertion in the promoter region. This double mutant expressed the malPQ operon exactly as the wild-type strain did, in a maltose-inducible manner. In this strain, the starting point for malPQ transcription was shifted 11 nucleotides downstream from the wild-type location. An analysis of these results suggests that (i) the binding site for the malT product is located upstream from the region which is severely altered in the double mutant, i.e., upstream from position -31; and (ii) the 30-base pair sequence which precedes the transcription starting point contains very few positions which are essential for promoter activity.
麦芽糖操纵子malPQ是麦芽糖调节子的三个操纵子之一,受基因malT产物的正调控。malPQ转录的起始点是通过一系列实验推导出来的,这些实验包括将体内合成的malPQ mRNA与合适的DNA探针杂交,然后要么消化未杂交的DNA(S1核酸酶图谱分析),要么延伸杂交探针(逆转录酶图谱分析)。在野生型菌株中,这个起始点位于malP起始密码子上游37个核苷酸处。对一个双突变体也进行了这种分析,该双突变体在启动子区域既有一个13碱基对的缺失,又有一个3碱基对的插入。这个双突变体以麦芽糖诱导的方式表达malPQ操纵子,与野生型菌株完全一样。在这个菌株中,malPQ转录的起始点从野生型位置向下游移动了11个核苷酸。对这些结果的分析表明:(i)malT产物的结合位点位于双突变体中严重改变的区域上游,即-31位上游;(ii)转录起始点之前的30碱基对序列中,对启动子活性至关重要的位置很少。