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首页> 美国卫生研究院文献>Applied and Environmental Microbiology >DNA Microarray Analysis of Nitrogen Fixation and Fe(III) Reduction in Geobacter sulfurreducens
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DNA Microarray Analysis of Nitrogen Fixation and Fe(III) Reduction in Geobacter sulfurreducens

机译：DNA芯片分析减少固硫菌中固氮和还原Fe（III）的能力

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摘要

A DNA microarray representing the genome of Geobacter sulfurreducens was constructed for use in global gene expression profiling of cells under steady-state conditions with acetate as the electron donor and Fe(III) or fumarate as the electron acceptor. Reproducible differences in transcript levels were also observed in comparisons between cells grown with ammonia and those fixing atmospheric nitrogen. There was a high correlation between changes in transcript levels determined with microarray analyses and an evaluation of a subset of the genome with quantitative PCR. As expected, cells required to fix nitrogen had higher levels of transcripts of genes associated with nitrogen fixation, further demonstrating that the microarray approach could reliably detect important physiological changes. Cells grown with Fe(III) as the electron acceptor had higher levels of transcripts for omcB, a gene coding for an outer membrane c-type cytochrome that is essential for Fe(III) reduction. Several other c-type cytochrome genes also appeared to be up-regulated. An unexpected result was significantly higher levels of transcripts for genes which have a role in metal efflux, potentially suggesting the importance of maintaining metal homeostasis during release of soluble metals when reducing Fe(III). A substantial proportion (30%) of significantly expressed genes during Fe(III) reduction were genes of unknown function or hypothetical proteins, suggesting differences in Fe(III) reduction physiology among microorganisms which perform this metabolic process.
机译：构建了代表减少硫杆菌的基因组的DNA微阵列，用于在稳态条件下以乙酸盐作为电子供体，Fe（III）或富马酸盐作为电子受体的细胞在全局基因表达谱中的应用。在用氨气生长的细胞与固定大气氮的细胞之间进行比较时，也观察到了转录水平的可再现差异。通过微阵列分析确定的转录水平变化与通过定量PCR评估基因组子集之间存在高度相关性。正如预期的那样，固定氮所需的细胞具有更高水平的与固氮相关的基因转录本，进一步证明了微阵列方法可以可靠地检测重要的生理变化。以Fe（III）作为电子受体生长的细胞具有更高水平的omcB转录本，omcB是编码外膜c型细胞色素的基因，该基因对于Fe（III）还原至关重要。其他几个c型细胞色素基因也似乎被上调。出乎意料的结果是，具有在金属外排中起作用的基因的转录物水平显着升高，这可能表明在还原Fe（III）时，在可溶性金属释放过程中保持金属稳态的重要性。 Fe（III）还原过程中显着表达的基因中有很大一部分（30％）是功能未知或假设蛋白的基因，这表明执行该代谢过程的微生物之间Fe（III）还原生理的差异。

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