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表达伯克霍尔德氏菌 xenovorans LB400 的六种铬酸盐离子转运体同源物。

Expression of the six chromate ion transporter homologues of Burkholderia xenovorans LB400.

机构信息

Instituto de Investigaciones Químico-Biológicas, Universidad Michoacana, Morelia, Michoacán, Mexico.

Laboratorio Estatal de Salud Pública, Secretaría de Salud de Michoacán, Morelia, Michoacán, Mexico.

出版信息

Microbiology (Reading). 2014 Feb;160(Pt 2):287-295. doi: 10.1099/mic.0.073783-0. Epub 2013 Nov 20.

Abstract

The chromate ion transporter (CHR) superfamily comprises transporters that confer chromate resistance by extruding toxic chromate ions from cytoplasm. Burkholderia xenovorans strain LB400 has been reported to encode six CHR homologues in its multireplicon genome. We found that strain LB400 displays chromate-inducible resistance to chromate. Susceptibility tests of Escherichia coli strains transformed with cloned B. xenovorans chr genes indicated that the six genes confer chromate resistance, although under different growth conditions, and suggested that expression of chr genes is regulated by sulfate. Expression of chr genes was measured by quantitative reverse transcription-PCR (RT-qPCR) from total RNA of B. xenovorans LB400 grown under different concentrations of sulfate and exposed or not to chromate. The chr homologues displayed distinct expression levels, but showed no significant differences in transcription under the various sulfate concentrations tested, indicating that sulfate does not regulate chr gene expression in B. xenovorans. The chrA2 gene, encoded in the megaplasmid, was the only chr gene whose expression was induced by chromate and it was shown to constitute the chromate-responsive chrBACF operon. These data suggest that this determinant is mainly responsible for the B. xenovorans LB400 chromate resistance phenotype.

摘要

铬酸盐转运蛋白(CHR)超家族包括通过将有毒的铬酸盐离子从细胞质中排出而赋予铬酸盐抗性的转运蛋白。据报道,伯克霍尔德氏菌属 Xenovorans 菌株 LB400 在其多复制子基因组中编码六个 CHR 同源物。我们发现,菌株 LB400 对铬酸盐表现出可诱导的抗性。用克隆的 B. xenovorans chr 基因转化的大肠杆菌菌株的敏感性测试表明,这六个基因赋予了铬酸盐抗性,尽管在不同的生长条件下,并且表明 chr 基因的表达受硫酸盐调节。通过定量逆转录 PCR(RT-qPCR)从在不同硫酸盐浓度下生长的 B. xenovorans LB400 的总 RNA 测量 chr 基因的表达,并暴露或不暴露于铬酸盐。chr 同源物显示出不同的表达水平,但在测试的各种硫酸盐浓度下转录没有显着差异,表明硫酸盐在 B. xenovorans 中不调节 chr 基因表达。在大质粒中编码的 chrA2 基因是唯一被铬酸盐诱导表达的 chr 基因,它构成了铬酸盐响应的 chrBACF 操纵子。这些数据表明,该决定因素主要负责 B. xenovorans LB400 的铬酸盐抗性表型。

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