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转录衰减及mRNA稳定性差异在大肠杆菌蜜二糖操纵子调控中的作用

Transcriptional attenuation and differential mRNA stability in the regulation of the Escherichia coli melibiose operon.

作者信息

Shimamoto T, Noguchi K, Kuroda M, Tsuda M, Tsuchiya T

机构信息

Gene Research Center, Faculty of Pharmaceutical Sciences, Okayama University.

出版信息

J Biochem. 1994 Jun;115(6):1185-9. doi: 10.1093/oxfordjournals.jbchem.a124477.

Abstract

The organization of the melibiose operon of Escherichia coli is promoter-melA-melB. The amount of the product (alpha-galactosidase) of the first gene (melA) is much larger than that of the product (melibiose permease) of the second gene (melB). Using the chloramphenicol acetyltransferase gene (cat) as reporter, we found that there was an element between melA and melB, which reduced the expression of the downstream gene, melB. This region contained a boxA-like sequence, which is known as a binding site for an attenuation factor, NusA. Northern hybridization analysis revealed that the ratio of melA mRNA and melAB mRNA was comparable with the ratio of the melA and melB products. We also found that the melA mRNA was about 3-fold more stable than the melAB mRNA. Experimental results obtained with a nusAts mutant suggested that the NusA protein is involved in the reduced expression of the melB gene. We conclude that the production ratio of alpha-galactosidase and melibiose permease is regulated at two levels: 1) transcription and 2) mRNA stability.

摘要

大肠杆菌蜜二糖操纵子的结构为启动子 - melA - melB。第一个基因(melA)的产物(α - 半乳糖苷酶)的量远大于第二个基因(melB)的产物(蜜二糖通透酶)的量。以氯霉素乙酰转移酶基因(cat)作为报告基因,我们发现在melA和melB之间存在一个元件,它降低了下游基因melB的表达。该区域包含一个类似boxA的序列,它是衰减因子NusA的结合位点。Northern杂交分析表明,melA mRNA与melAB mRNA的比例与melA和melB产物的比例相当。我们还发现melA mRNA的稳定性比melAB mRNA高约3倍。用nusA温度敏感突变体获得的实验结果表明,NusA蛋白参与了melB基因表达的降低。我们得出结论,α - 半乳糖苷酶和蜜二糖通透酶的产生比例在两个水平上受到调控:1)转录;2)mRNA稳定性。

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