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不同氮素可利用条件下马氏甲烷八叠球菌菌株Gö1的全转录组分析

Global transcriptional analysis of Methanosarcina mazei strain Gö1 under different nitrogen availabilities.

作者信息

Veit Katharina, Ehlers Claudia, Ehrenreich Armin, Salmon Kirsty, Hovey Raymond, Gunsalus Robert P, Deppenmeier Uwe, Schmitz Ruth A

机构信息

Institut für Allgemeine Mikrobiologie, Christian-Albrechts-Universität Kiel, Am Botanischen Garten 1-9, 24118, Kiel, Germany.

出版信息

Mol Genet Genomics. 2006 Jul;276(1):41-55. doi: 10.1007/s00438-006-0117-9. Epub 2006 Apr 20.

Abstract

Certain archaeal species can fix molecular nitrogen under nitrogen limiting conditions although little is known about this process at either the genetic or molecular level. To address this on a genome-wide scale, transcriptional analysis was performed on the model methanogen Methanosarcina mazei strain Gö1 using DNA-microarrays. The genomic expression patterns for cells grown under nitrogen fixing conditions versus nitrogen sufficiency (10 mM ammonium) revealed that approximately 5% of all genes are differentially expressed. Besides a small set of genes previously known to be up-regulated under nitrogen limitation, 14 additional genes involved in nitrogen metabolism were identified plus 10 genes encoding potential transcriptional regulators, 13 genes involved in carbon metabolism, 3 genes in general stress response, 8 putative transporter genes, and an additional 21 genes with unknown function. Quantitative reverse transcriptase PCR experiments confirmed the differential expression of a subset of these genes. Promoter analysis revealed a palindromic DNA motif centered nearby the transcriptional start point for several genes up-regulated under nitrogen limitation. A bioinformatics study demonstrated the presence of this motif in the up-stream region of 52 genes genome-wide, the majority of which showed nitrogen dependent differential transcription. We therefore hypothesize that this DNA element is involved in nitrogen control in M. mazei where it may act as a binding site for a regulatory protein.

摘要

某些古细菌物种能够在氮限制条件下固定分子氮,尽管在遗传或分子水平上对这一过程了解甚少。为了在全基因组范围内解决这个问题,我们使用DNA微阵列对模式产甲烷菌马氏甲烷八叠球菌菌株Gö1进行了转录分析。在固氮条件下生长的细胞与氮充足条件(10 mM铵)下生长的细胞的基因组表达模式显示,所有基因中约5%存在差异表达。除了一小部分先前已知在氮限制下上调的基因外,还鉴定出14个参与氮代谢的额外基因,加上10个编码潜在转录调节因子的基因、13个参与碳代谢的基因、3个参与一般应激反应的基因、8个推定的转运蛋白基因,以及另外21个功能未知的基因。定量逆转录酶PCR实验证实了这些基因子集中的差异表达。启动子分析揭示了一个回文DNA基序,位于几个在氮限制下上调的基因转录起始点附近。一项生物信息学研究表明,在全基因组范围内,52个基因的上游区域存在这种基序,其中大多数显示出氮依赖性差异转录。因此,我们假设这个DNA元件参与了马氏甲烷八叠球菌的氮控制,它可能作为一种调节蛋白的结合位点。

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