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编码大肠杆菌主要外膜蛋白的ompA基因启动子的特性分析。

Characterisation of the promoters for the ompA gene which encodes a major outer membrane protein of Escherichia coli.

作者信息

Cole S T, Bremer E, Hindennach I, Henning U

出版信息

Mol Gen Genet. 1982;188(3):472-9. doi: 10.1007/BF00330051.

Abstract

The regulatory region of the ompA gene from Escherichia coli has been characterized by biochemical and genetic approaches. Two overlapping promoters, P1 and P2, organized in that order with respect to the ompA coding sequence, were identified and it was found that ompA possesses an unusually long leader region. Both P1 and P2 were active in an in vitro transcription system although S1 mapping analysis of the ompA mRNA made in vivo showed that P2 was mainly responsible for transcription of the gene. Confirmation of this was obtained by studying down-promoter mutants of ompA cloned in pSC101. These mutants were classified into two groups, deletions and insertions. The deletions, which were caused by the IS102 insertion element found in pSC101 removed the--35 regions of both P1 and P2. However, since P2 was distally situated with respect to the IS element it was less extensively damaged and it is proposed that the residual P2 sequence is responsible for the low level of expression observed. In addition to an IS102 insertion in the promoter region four IS1 insertion mutants were characterized. These had integrated at different positions in the ompA leader region and were all incompletely polar.

摘要

利用生化和遗传学方法对大肠杆菌ompA基因的调控区进行了表征。鉴定出两个重叠的启动子P1和P2,它们相对于ompA编码序列按此顺序排列,并且发现ompA具有异常长的前导区。P1和P2在体外转录系统中均有活性,尽管对体内产生的ompA mRNA进行的S1图谱分析表明P2主要负责该基因的转录。通过研究克隆在pSC101中的ompA的启动子下游突变体得到了这一点的证实。这些突变体分为两组,即缺失突变体和插入突变体。由pSC101中发现的IS102插入元件引起的缺失突变去除了P1和P2的-35区域。然而,由于P2相对于IS元件位于远端,其受损程度较小,因此有人提出残留的P2序列是观察到的低水平表达的原因。除了在启动子区域有一个IS102插入外,还对四个IS1插入突变体进行了表征。它们整合在ompA前导区的不同位置,并且都不是完全极性的。

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