枯草芽孢杆菌色氨酸启动子区域的特性分析
Characterization of the Bacillus subtilis tryptophan promoter region.
作者信息
Shimotsu H, Henner D J
出版信息
Proc Natl Acad Sci U S A. 1984 Oct;81(20):6315-9. doi: 10.1073/pnas.81.20.6315.
Abstract
The nucleotide sequence of the control region of the trp operon of Bacillus subtilis has been determined. The region was shown to contain the trp promoter by deletion analysis and by determination of the transcription start site. The trp promoter shows similarity to the consensus sequence for Escherichia coli and B. subtilis promoters. The presence of the trp control region on a high-copy-number plasmid confers resistance to the tryptophan analogue 5-methyltryptophan. It appears that an approximately 120-base-pair region comprising not only the trp promoter but also adjacent direct repeat sequences is necessary to confer 5-methyltryptophan resistance. We postulate that this region is involved in tryptophan regulation and confers 5-methyltryptophan resistance by titration of a trp regulatory protein. Removal of either the trp promoter or the adjacent direct repeat sequences abolished the 5-methyltryptophan-resistance phenotype. Placement of unrelated promoters adjacent to the direct repeat sequences restored 5-methyltryptophan resistance. This suggests that promoter activity is necessary for the regulatory function.
摘要
枯草芽孢杆菌色氨酸操纵子调控区的核苷酸序列已被确定。通过缺失分析和转录起始位点的测定表明，该区域包含色氨酸启动子。色氨酸启动子与大肠杆菌和枯草芽孢杆菌启动子的共有序列相似。高拷贝数质粒上存在色氨酸调控区可赋予对色氨酸类似物5-甲基色氨酸的抗性。似乎一个约120个碱基对的区域，不仅包括色氨酸启动子，还包括相邻的直接重复序列，对于赋予5-甲基色氨酸抗性是必需的。我们推测该区域参与色氨酸调控，并通过滴定一种色氨酸调节蛋白赋予5-甲基色氨酸抗性。去除色氨酸启动子或相邻的直接重复序列会消除5-甲基色氨酸抗性表型。将无关启动子置于直接重复序列附近可恢复5-甲基色氨酸抗性。这表明启动子活性对于调控功能是必需的。
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