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编码来自绿针假单胞菌O6的有机酸转运蛋白的dctA基因的转录调控和突变分析

Transcriptional regulation and mutational analysis of a dctA gene encoding an organic acid transporter protein from Pseudomonas chlororaphis O6.

作者信息

Nam Hyo Song, Spencer Matt, Anderson Anne J, Cho Baik Ho, Kim Young Cheol

机构信息

Agricultural Plant Stress Research Center and Inst. of Agri. Sci. and Tech., College of Agriculture and Life Sciences, Chonnam National University, Gwangju 500-757, South Korea.

出版信息

Gene. 2003 Dec 24;323:125-31. doi: 10.1016/j.gene.2003.09.012.

Abstract

A dctA gene encoding a protein with identity to a C(4)-dicarboxylic acid/H(+) symporter was cloned from a beneficial root colonizer, Pseudomonas chlororaphis O6 (PcO6). Expression of the dctA gene was induced in minimal medium by several organic acids and was repressed by glucose. Highest expression was observed in early-logarithmic (log) cells grown on fumarate, acetate or succinate with decline as cells approached late-log growth phase. The dctA transcript accumulated weakly when cells were grown on malate, but strong expression was observed with benzoate. Expression of the dctA transcript was repressed in early-log cells upon addition of glucose to fumarate, but was detected as the cell culture aged. A dctA-deficient mutant of PcO6, constructed by marker exchange mutagenesis, did not grow on minimal medium containing succinate, benzoate, acetate or fumarate and growth on malate was delayed. The dctA mutant and wild-type grew equally on citrate, glucose, fructose, sucrose or inositol. We conclude that the transporter protein encoded by dctA is essential for utilization of certain organic acids and its expression is controlled by the availability of sugars.

摘要

从有益的根定殖菌——绿针假单胞菌O6(PcO6)中克隆到一个dctA基因,该基因编码的蛋白质与C(4)-二羧酸/H(+)同向转运体具有同源性。dctA基因的表达在基本培养基中被几种有机酸诱导,并被葡萄糖抑制。在以富马酸、乙酸或琥珀酸为碳源生长的对数早期(log)细胞中观察到最高表达,随着细胞接近对数生长后期表达下降。当细胞以苹果酸为碳源生长时,dctA转录本积累较弱,但以苯甲酸为碳源时观察到强表达。在以富马酸为碳源的对数早期细胞中添加葡萄糖后,dctA转录本的表达受到抑制,但随着细胞培养时间延长可检测到表达。通过标记交换诱变构建的PcO6的dctA缺陷突变体,在含有琥珀酸、苯甲酸、乙酸或富马酸的基本培养基上不能生长,在以苹果酸为碳源时生长延迟。dctA突变体和野生型在柠檬酸盐、葡萄糖、果糖、蔗糖或肌醇上生长情况相同。我们得出结论,dctA编码的转运蛋白对于某些有机酸的利用至关重要,其表达受糖的可用性控制。

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