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奇异变形杆菌中大肠杆菌乳糖操纵子的异常表达。I. L8和L8 UV5的作用。

Anomalous expression of the E. coli lac operon in Proteus mirabilis. I. Effects of L8 and L8 UV5.

作者信息

Roberts M, Baumberg S

出版信息

Mol Gen Genet. 1984;198(2):159-65. doi: 10.1007/BF00328716.

Abstract

The lac operon shows anomalous expression in Proteus mirabilis: the maximal induced level is 10% or less of that in E. coli, while repression reduces this by a factor of only 2-5. We have sought to determine whether this effect relates in any way to CRP-mediated activation of expression, by comparing expression in P. mirabilis of lac operons (introduced for technical reasons on IncP1 plasmids) either regulatorily wild-type or bearing L8 or L8UV5. Derivatives of RP1 bearing L8UV5 were obtained by homogenotisation of pGC9114 (RP1::Tn951) in a L8UV5 background; while derivatives of RP4 bearing lac+, L8 or L8UV5 were obtained by Mu-mediated translocation of chromosomal regions bearing these alleles, following partial heat-induction of Mucts62 on pGM14 (RP4::Mucts62) in the appropriate hosts. These plasmids could be readily transferred to, and stably maintained in, the P. mirabilis strains employed. It was found that L8 reduced the maximal level of beta-galactosidase activity, and L8UV5 restored this activity to around wild-type, in P. mirabilis quantitatively very much as in E. coli. Nevertheless, the low maximal level of expression and high basal level characteristic of the former host were unchanged. The simplest explanation of these results is that P. mirabilis contains a protein that mimics the E. coli CRP protein in interacting with the appropriate DNA binding site and thereby stimulating transcription; and that the anomalous regulation of lac in this host is unconnected with the CRP system.

摘要

乳糖操纵子在奇异变形杆菌中表现出异常表达

最大诱导水平仅为大肠杆菌中的10%或更低,而阻遏作用仅使其降低2 - 5倍。我们试图通过比较奇异变形杆菌中调控野生型或携带L8或L8UV5的乳糖操纵子(因技术原因引入IncP1质粒)的表达,来确定这种效应是否以任何方式与CRP介导的表达激活有关。携带L8UV5的RP1衍生物是通过在L8UV5背景下对pGC9114(RP1::Tn951)进行同源重组获得的;而携带lac +、L8或L8UV5的RP4衍生物是通过在适当宿主中对pGM14(RP4::Mucts62)上的Mucts62进行部分热诱导后,Mu介导的携带这些等位基因的染色体区域易位获得的。这些质粒可以很容易地转移到所用的奇异变形杆菌菌株中并稳定维持。结果发现,在奇异变形杆菌中,L8降低了β - 半乳糖苷酶活性的最大水平,而L8UV5将该活性恢复到接近野生型水平,在数量上与在大肠杆菌中非常相似。然而,前一种宿主中低的最大表达水平和高的基础水平特征并未改变。对这些结果最简单的解释是,奇异变形杆菌含有一种蛋白质,它在与适当的DNA结合位点相互作用并由此刺激转录方面模仿了大肠杆菌的CRP蛋白;并且该宿主中乳糖操纵子的异常调控与CRP系统无关。

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