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鼠伤寒沙门氏菌腺苷酸环化酶基因(cya)表达与调控重要序列元件的分析

Analysis of sequence elements important for expression and regulation of the adenylate cyclase gene (cya) of Salmonella typhimurium.

作者信息

Thorner L K, Fandl J P, Artz S W

机构信息

Department of Microbiology, University of California, Davis 95616.

出版信息

Genetics. 1990 Aug;125(4):709-17. doi: 10.1093/genetics/125.4.709.

Abstract

We determined the nucleotide sequence of the regulatory region of the cya gene of Salmonella typhimurium. A set of nested BAL-31 deletions originating upstream of the promoter/regulatory region and extending into the cya structural gene was constructed in M13mp::cya phages and was tested for complementation of a chromosomal cya deletion mutation. BAL-31 deletion mutants unable to complement cya localized the major cya promoter. The synthetic tac promoter was inserted upstream of the BAL-31 deletions so that expression of cya was dependent on transcription from tac. Those tac derivative phages unable to complement cya localized the translation initiation region. The cya DNA sequence revealed at least three potential promoters capable of transcribing cya, with a CRP binding site straddling the-10 hexamer of the promoter proximal to the structural gene. The leader RNA sequence initiated at the latter promoter is approximately 140 bases long and includes a region that may form a stable secondary structure (delta G = -23.8 kcal). There exist two possible in-frame translation start points, one of which is TTG and the other of which is ATG. The sequence of the S. typhimurium regulatory region was compared with that reported for Escherichia coli.

摘要

我们测定了鼠伤寒沙门氏菌cya基因调控区的核苷酸序列。在M13mp::cya噬菌体中构建了一组从启动子/调控区上游起始并延伸至cya结构基因的嵌套BAL-31缺失体,并检测其对染色体cya缺失突变的互补作用。无法互补cya的BAL-31缺失突变体定位了主要的cya启动子。将合成的tac启动子插入到BAL-31缺失体的上游,使得cya的表达依赖于tac的转录。那些无法互补cya的tac衍生噬菌体定位了翻译起始区。cya的DNA序列显示至少有三个能够转录cya的潜在启动子,一个CRP结合位点横跨结构基因近端启动子的-10六聚体。从后一个启动子起始的前导RNA序列约140个碱基长,并且包括一个可能形成稳定二级结构(ΔG = -23.8千卡)的区域。存在两个可能的读框内翻译起始点,其中一个是TTG,另一个是ATG。将鼠伤寒沙门氏菌调控区的序列与已报道的大肠杆菌的序列进行了比较。

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本文引用的文献

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