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Desulfovibrio sp. Genes Involved in the Respiration of Sulfate during Metabolism of Hydrogen and Lactate

机译:脱硫弧菌氢和乳酸代谢过程中涉及硫酸盐呼吸的基因

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摘要

To develop a better understanding of respiration by sulfate-reducing bacteria, we examined transcriptional control of respiratory genes during growth with lactate or hydrogen as an electron donor. RNA extracts of Desulfovibrio desulfuricans subsp. aestuarii were analyzed by using random arbitrarily primed PCR. RNA was reverse transcribed under low-stringency conditions with a set of random primers, and candidate cDNAs were cloned, sequenced, and characterized by BLAST analysis. Putative differentially expressed transcripts were confirmed by Northern blot analysis. Interestingly, dissimilatory bisulfite reductase was upregulated in the presence of hydrogen. To link these transcriptional changes to the physiology of sulfate-reducing bacteria, sulfide was measured during growth of several strains of Desulfovibrio on hydrogen or lactate, and this revealed that hydrogen-grown cells produced more sulfide per unit of cell mass than lactate-grown cells. Transcription of other redox proteins was characterized by Northern blotting to determine whether or not they were also transcribed to higher levels in hydrogen-grown cells. Growth on lactate produced greater transcription of [NiFe] hydrogenase. H2-grown cells transcribed the adenylylsulfate reductase b subunit and HmcA to higher levels. The results we describe here provide new insight into the continuing debate over how Desulfovibrio species utilize redox components to generate membrane potential and to channel electrons to sulfate, the final electron acceptor.
机译:为了更好地理解减少硫酸盐的细菌的呼吸作用,我们在乳酸或氢作为电子供体的生长过程中检查了呼吸基因的转录控制。 Desulfovibrio desulfuricans亚种的RNA提取物。通过使用随机任意引发的PCR分析河口。在低严格条件下,用一组随机引物对RNA进行反转录,并通过BLAST分析对候选cDNA进行克隆,测序和鉴定。通过Northern印迹分析确认推定的差异表达的转录物。有趣的是,在氢存在下,异化亚硫酸氢盐还原酶被上调。为了将这些转录变化与减少硫酸盐的细菌的生理联系起来,在数种脱硫弧菌菌株在氢或乳酸上生长期间测量了硫化物,这表明氢生长的细胞比乳酸生长的细胞每单位细胞质量产生更多的硫化物。 。通过Northern印迹表征其他氧化还原蛋白的转录,以确定它们是否在氢生长细胞中也转录到更高的水平。在乳酸上的生长产生[NiFe]氢化酶更大的转录。 H2生长的细胞将腺苷酸硫酸还原酶b亚基和HmcA转录到更高的水平。我们在此描述的结果为有关Desulfovibrio物种如何利用氧化还原成分产生膜电位并将电子引导至最终的电子受体硫酸盐的持续辩论提供了新的见识。

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