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枯草芽孢杆菌L-阿拉伯糖(ara)操纵子:核苷酸序列、基因组织与表达

The Bacillus subtilis L-arabinose (ara) operon: nucleotide sequence, genetic organization and expression.

作者信息

Sa-Nogueira Isabel, Nogueira Teresa V, Soares Snia, de Lencastre Hermnia

机构信息

Instituto de Tecnologia Química e Biológica Universidade Nova de Lisboa. Apartado 127, 2780 Oeiras Codex Portugal.

The Rockefeller UniversityLaboratory of Microbiology 1230 York Avenue, New York, NY 10021-6399 USA.

出版信息

Microbiology (Reading). 1997 Mar;143 ( Pt 3):957-969. doi: 10.1099/00221287-143-3-957.

DOI:10.1099/00221287-143-3-957
PMID:9084180
Abstract

The Bacillus subtilis L-arabinose metabolic genes araA, araB and araD, encoding L-arabinose isomerase, L-ribulokinase and L-ribulose-5-phosphate 4-epimerase, respectively, have been cloned previously and the products of araB and araD were shown to be functionally homologous to their Escherichia coli counterparts by complementation experiments. Here we report that araA, araB and araD, whose inactivation leads to an Ara- phenotype, are the first three ORFs of a nine cistron transcriptional unit with a total length of 11 kb. This operon, called ara, is located at about 256 degrees on the B. subtilis genetic map and contains six new genes named araL, araM, araN, araP, araQ and abfA. Expression of the ara operon is directed by a strong sigma A-like promoter identified within a 150 bp DNA fragment upstream from the translation start site of araA. Analysis of the sequence of the ara operon showed that the putative products of araN, araP and araQ are homologous to bacterial components of binding-protein-dependent transport systems and abfA most probably encodes an alpha-L-arabinofuranosidase. The functions of araL and araM are unknown. An in vitro-constructed insertion-deletion mutation in the region downstream from araD allowed us to demonstrate that araL, araM, araN, araP, araQ and abfA are not essential for L-arabinose utilization. Studies with strains bearing transcriptional fusions of the operon to the E. coli lacZ gene revealed that expression from the ara promoter is induced by L-arabinose and repressed by glucose.

摘要

枯草芽孢杆菌的L-阿拉伯糖代谢基因araA、araB和araD,分别编码L-阿拉伯糖异构酶、L-核酮糖激酶和L-核酮糖-5-磷酸4-差向异构酶,此前已被克隆,并且通过互补实验表明araB和araD的产物与其大肠杆菌对应物在功能上具有同源性。在此我们报告,其失活导致Ara-表型的araA、araB和araD是一个全长11 kb的九个顺反子转录单元的前三个开放阅读框。这个操纵子称为ara,位于枯草芽孢杆菌遗传图谱上约256度处,包含六个新基因,分别命名为araL、araM、araN、araP、araQ和abfA。ara操纵子的表达由一个在araA翻译起始位点上游150 bp DNA片段内鉴定出的强σA样启动子指导。对ara操纵子序列的分析表明,araN、araP和araQ的推定产物与依赖结合蛋白的转运系统的细菌组分同源,而abfA很可能编码一种α-L-阿拉伯呋喃糖苷酶。araL和araM的功能未知。在araD下游区域体外构建的插入缺失突变使我们能够证明araL、araM、araN、araP、araQ和abfA对于L-阿拉伯糖的利用不是必需的。对带有该操纵子与大肠杆菌lacZ基因转录融合的菌株的研究表明,ara启动子的表达受L-阿拉伯糖诱导并受葡萄糖抑制。

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