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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Gene function analysis in environmental isolates: The nif regulon of the strict iron oxidizing bacterium Leptospirillum ferrooxidans
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Gene function analysis in environmental isolates: The nif regulon of the strict iron oxidizing bacterium Leptospirillum ferrooxidans

机译：环境分离物中的基因功能分析：严格的铁氧化细菌氧化亚铁螺旋体

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A random genomic library from an environmental isolate of the Gram-negative bacterium Leptospirillum ferrooxidans has been printed on a microarray. Gene expression analysis was carried out with total RNA extracted from L. ferrooxidans cultures in the presence or absence of ammonium as nitrogen source under aerobic conditions. Although practically nothing is known about the genome sequence of this bacterium, this approach allowed us the selection and sequencing of only those clones bearing genes that showed an altered expression pattern. By sequence comparison, we have identified most of the genes of nitrogen fixation regulon in L. ferrooxidans, like the nifHDKENX operon, encoding the structural components of Mo-Fe nitrogenase; nifSU-hesB-hscBA-fdx operon, for Fe-S cluster assembly; the amtB gene (ammonium transporter); modA (molybdenum ABC type transporter); some regulatory genes like ntrC, nifA (the specific activator of nif genes); or two glnB-like genes (encoding the PII regulatory protein). Our results show that shotgun DNA microarrays are very powerful tools to accomplish gene expression studies with environmental bacteria whose genome sequence is still unknown, avoiding the time and effort necessary for whole genome sequencing projects. [References: 40]
机译：来自革兰氏阴性细菌铁氧体螺旋体的环境分离株的随机基因组文库已印在微阵列上。在有氧条件下，在有或没有铵作为氮源的情况下，用从氧化亚铁乳酸杆菌培养物中提取的总RNA进行基因表达分析。尽管实际上对该细菌的基因组序列一无所知，但这种方法使我们仅能选择和测序带有表达模式改变基因的克隆。通过序列比较，我们已经确定了氧化亚铁乳酸菌中固氮调节子的大部分基因，如nifHDKENX操纵子，编码Mo-Fe固氮酶的结构成分。 nifSU-hesB-hscBA-fdx操纵子，用于Fe-S簇组装； amtB基因（铵转运蛋白）； modA（钼ABC型转运蛋白）；一些调节基因，例如ntrC，nifA（nif基因的特定激活物）；或两个glnB样基因（编码PII调节蛋白）。我们的结果表明，shot弹枪DNA微阵列是非常有效的工具，可用于对环境细菌进行基因表达研究，而环境细菌的基因组序列仍然未知，从而避免了整个基因组测序项目所需的时间和精力。 [参考：40]

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