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苜蓿中华根瘤菌中用于赤藓糖醇转运和分解代谢的必需基因座。

A locus necessary for the transport and catabolism of erythritol in Sinorhizobium meliloti.

机构信息

Department of Microbiology, University of Manitoba, Winnipeg, MB R3T 2N2, Canada.

出版信息

Microbiology (Reading). 2010 Oct;156(Pt 10):2970-2981. doi: 10.1099/mic.0.041905-0. Epub 2010 Jul 29.

Abstract

In this work we have genetically defined an erythritol utilization locus in Sinorhizobium meliloti. A cosmid containing the locus was isolated by complementation of a transposon mutant and was subsequently mutagenized using Tn5 : : B20. The locus was found to consist of five transcriptional units, each of which was necessary for the utilization of erythritol. Genetic complementation experiments using genes putatively annotated as erythritol catabolic genes clearly showed that, of the 17 genes at this locus, six genes are not necessary for the utilization of erythritol as a sole carbon source. The remaining genes encode EryA, EryB, EryC and TpiB as well as an uncharacterized ABC-type transporter. Transport experiments using labelled erythritol showed that components of the ABC transporter are necessary for the uptake of erythritol. The locus also contains two regulators: EryD, a SorC class regulator, and SMc01615, a DeoR class regulator. Quantitative RT-PCR experiments showed that each of these regulators negatively regulates its own transcription. In addition, induction of the erythritol locus was dependent upon EryD and a product of erythritol catabolism. Further characterization of polar mutations revealed that in addition to erythritol, the locus contains determinants for adonitol and l-arabitol utilization. The context of the mutations suggests that the locus is important for both the transport and catabolism of adonitol and l-arabitol.

摘要

在这项工作中,我们在中华根瘤菌中遗传定义了赤藓糖醇利用基因座。通过转座子突变体的互补性分离出含有该基因座的柯斯质粒,并随后使用 Tn5∶B20 对其进行诱变。该基因座由五个转录单元组成,每个单元都需要赤藓糖醇的利用。使用推定注释为赤藓糖醇分解代谢基因的基因进行遗传互补实验清楚地表明,在这个基因座的 17 个基因中,有 6 个基因不是利用赤藓糖醇作为唯一碳源所必需的。其余基因编码 EryA、EryB、EryC 和 TpiB 以及一个未鉴定的 ABC 型转运蛋白。使用标记的赤藓糖醇进行的运输实验表明,ABC 转运蛋白的成分是赤藓糖醇摄取所必需的。该基因座还包含两个调节剂:EryD,一种 SorC 类调节剂,和 SMc01615,一种 DeoR 类调节剂。定量 RT-PCR 实验表明,这些调节剂中的每一种都负调控其自身的转录。此外,赤藓糖醇基因座的诱导依赖于 EryD 和赤藓糖醇分解代谢产物。对极性突变的进一步表征表明,除了赤藓糖醇外,该基因座还包含阿东醇和 L-阿拉伯糖醇利用的决定因素。突变的上下文表明,该基因座对阿东醇和 L-阿拉伯糖醇的运输和分解代谢都很重要。

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