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Expression of the Escherichia coli malPQ operon remains unaffected after drastic alteration of its promoter.

机译:大肠杆菌malPQ操纵子的表达在其启动子发生巨大变化后仍然不受影响。

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摘要

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.
机译:麦芽糖调节子的三个操纵子之一malPQ操纵子受malT基因产物的正调控。 malPQ转录的起点是从以下实验得出的,该实验涉及体内合成的malPQ mRNA与适当的DNA探针杂交,然后消化未杂交的DNA(S1核酸酶作图)或杂交探针的延伸(逆转录酶)映射)。在野生型菌株中,该起点是malP起始密码子上游37个核苷酸。还对在启动子区域中既包含13个碱基对缺失又包含3个碱基对插入的双突变体进行了该分析。该双突变体以麦芽糖可诱导的方式与野生型菌株完全一样表达malPQ操纵子。在该菌株中,malPQ转录的起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的上游,即位置-31的上游; (ii)在转录起始点之前的30个碱基对的序列包含很少的位置,这些位置对于启动子活性是必不可少的。

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