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首页> 外文期刊>Journal of Molecular Biology >Differential Role of Base Pairs on gal Promoters Strength
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Differential Role of Base Pairs on gal Promoters Strength

机译：碱基对对gal启动子强度的差异作用

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摘要

Sequence alignments of promoters in prokaryotes postulated that the frequency of occurrence of a base pair at a given position of promoter elements reflects its contribution to intrinsic promoter strength. We directly assessed the contribution of the four base pairs in each position in the intrinsic promoter strength by keeping the context constant in Escherichia coil cAMP-CRP (cAMP receptor protein) regulated gal promoters by in vitro transcription assays. First, we show that base pair frequency within known consensus elements correlates well with promoter strength. Second, we observe some substitutions upstream of the ex-10 TG motif that are important for promoter function. Although the galP1 and P2 promoters overlap, only three positions where substitutions inactivated both promoters were found. We propose that RNA polymerase binds to the - 12T base pair as part of double-stranded DNA while opening base pairs from - 11A to +3 to form the single-stranded transcription bubble DNA during isomerization. The cAMP-CRP complex rescued some deleterious substitutions in the promoter region. The base pair roles and their flexibilities reported here for Escherichia coil gal promoters may help construction of synthetic promoters in gene circuitry experiments in which overlapping promoters with differential controls may be warranted. Published by Elsevier Ltd.
机译：推测原核生物中启动子的序列比对表明在启动子元件给定位置上碱基对的出现频率反映了其对固有启动子强度的贡献。我们通过体外转录测定法，通过保持大肠埃希氏线圈cAMP-CRP（cAMP受体蛋白）调节的gal启动子中的上下文恒定，直接评估了内源启动子强度中每个位置的四个碱基对的贡献。首先，我们表明已知共有元件内的碱基对频率与启动子强度密切相关。第二，我们观察到了ex-10 TG基序上游的一些替代，这些替代对于启动子功能很重要。尽管galP1和P2启动子重叠，但是仅发现三个位置使两个启动子失活。我们提出，RNA聚合酶作为双链DNA的一部分与-12T碱基对结合，同时从-11A到+3打开碱基对以在异构化过程中形成单链转录气泡DNA。 cAMP-CRP复合物挽救了启动子区域的一些有害取代。此处报道的大肠埃希氏菌线圈gal启动子的碱基对作用及其灵活性可能有助于构建基因电路实验中的合成启动子，在该实验中，可以保证重叠启动子具有不同的对照。由Elsevier Ltd.发布

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