Department of Civil and Environmental Engineering, University of California, 726 Davis Hall, Berkeley, CA 94720-1710, USA.
Biotechnol Bioeng. 2013 May;110(5):1333-41. doi: 10.1002/bit.24819. Epub 2013 Feb 9.
A microarray targeting four sequenced strains in the Dehalococcoides (Dhc) genus was used to analyze gene expression in a robust long-term trichloroethene (TCE)-degrading microbial community (designated ANAS) during feeding cycles that involve conditions of periodic substrate supply. The Dhc transcriptome was examined at three time-points throughout a batch feeding cycle: T1 (27 h) when TCE, dichloroethene (DCE), and vinyl chloride (VC) were present; T2 (54 h) when only VC remained; and T3 (13 days) when Dhc had been starved of substrate for 9 days. Ninety percent of the Dhc open reading frames (ORFs) that were detected in the ANAS DNA were found to be expressed as RNA sometime during the time course, demonstrating extraordinary utilization of the streamlined genome. Ninety-seven percent of these transcripts were differentially expressed during the time course indicating efficiency of transcription through regulation in Dhc. Most Dhc genes were significantly down-regulated at T3 , responding to a lack of substrate as would be expected. The tceA and vcrA genes, which code for proteins with known chlorinated ethene reduction functions, were highly expressed at both T1 and T2 , whereas two other putative reductive dehalogenase genes (DET0173 and DET1545) were most highly expressed at T2 , likely in response to the presence of VC. Hydrogenases were most highly expressed at T1 , reflecting their important role in accumulating electrons used to initiate reductive dechlorination and other biosynthesis pathways. Cobalamin transport genes were preferentially expressed at T2 , reflecting an increase in corrinoid transport as chloroethenes were degraded and a decrease in activity of the transport system after dehalogenation was complete. This is the first application of a microarray targeting a known genus, including both core genomes and identified strain-specific genes, to improve our understanding of transcriptional dynamics within an undefined microbial community.
使用针对 Dehalococcoides(Dhc)属中四个测序菌株的微阵列分析了在周期性底物供应条件下涉及周期性底物供应的强大长期三氯乙烯(TCE)降解微生物群落(指定为 ANAS)中的基因表达。在批处理循环的三个时间点检查了 Dhc 转录组:T1(27 h)时存在 TCE、二氯乙烯(DCE)和氯乙烯(VC);T2(54 h)时仅 VC 残留;T3(13 天)时 Dhc 已饥饿 9 天没有底物。在 ANAS DNA 中检测到的 90%的 Dhc 开放阅读框(ORF)在时间过程中某个时候被发现表达为 RNA,证明了对简化基因组的非凡利用。在时间过程中,这些转录本中有 97%差异表达,表明 Dhc 中的转录效率通过调节进行。大多数 Dhc 基因在 T3 时显著下调,这是对缺乏底物的预期反应。编码已知氯代乙稀还原功能蛋白的 tceA 和 vcrA 基因在 T1 和 T2 时均高度表达,而另外两个假定的还原脱卤酶基因(DET0173 和 DET1545)在 T2 时表达最高,可能是由于 VC 的存在。氢化酶在 T1 时表达最高,反映了它们在积累用于启动还原脱氯和其他生物合成途径的电子方面的重要作用。钴胺素转运基因在 T2 时优先表达,反映了随着氯代乙稀的降解,钴胺素转运的增加,以及脱卤化作用完成后转运系统活性的降低。这是首次将针对已知属的微阵列(包括核心基因组和鉴定的菌株特异性基因)应用于提高我们对未定义微生物群落中转录动态的理解。