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首页> 外文期刊>Applied and Environmental Microbiology >The Sequence of Spacers between the Consensus Sequences Modulates the Strength of Prokaryotic Promoters
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The Sequence of Spacers between the Consensus Sequences Modulates the Strength of Prokaryotic Promoters

机译：共有序列之间的间隔基序列调节原核启动子的强度

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We constructed a library of synthetic promoters forLactococcus lactis in which the known consensus sequences were kept constant while the sequences of the separating spacers were randomized. The library consists of 38 promoters which differ in strength from 0.3 up to more than 2,000 relative units, the latter among the strongest promoters known for this organism. The ranking of the promoter activities was somewhat different when assayed inEscherichia coli, but the promoters are efficient for modulating gene expression in this bacterium as well. DNA sequencing revealed that the weaker promoters (which had activities below 5 relative units) all had changes either in the consensus sequences or in the length of the spacer between the ?35 and ?10 sequences. The promoters in which those features were conserved had activities from 5 to 2,050 U, which shows that by randomizing the spacers, at least a 400-fold change in activity can be obtained. Interestingly, the entire range of promoter activities is covered in small steps of activity increase, which makes these promoters very suitable for quantitative physiological studies and for fine-tuning of gene expression in industrial bioreactors and cell factories.
机译：我们构建了乳酸乳球菌的合成启动子文库，其中已知的共有序列保持恒定，而分离间隔子的序列是随机的。该文库由38个启动子组成，这些启动子的强度从0.3到超过2,000个相对单位，后者是该生物已知的最强启动子。当在大肠杆菌中测定时，启动子活性的等级有所不同，但是启动子对于调节这种细菌中的基因表达也是有效的。 DNA测序表明，较弱的启动子（其活性低于5个相对单位）均在共有序列或？35和？10序列之间的间隔区长度上都有变化。保留了那些特征的启动子的活性为5至2050U，这表明通过使间隔子随机化，可以得到至少400倍的活性变化。有趣的是，启动子活性的整个范围都覆盖在增加活性的小步骤中，这使这些启动子非常适合于定量生理研究以及工业生物反应器和细胞工厂中基因表达的微调。

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