Re-annotation of the Saccharopolyspora erythraea genome using a systems biology approach.

Re-annotation of the Saccharopolyspora erythraea genome using a systems biology approach.
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DOI:
10.1186/1471-2164-14-699
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发表时间:
2013-10-11
期刊:
影响因子:
4.4
通讯作者:
Nielsen LK
Nielsen LK
中科院分区:
生物学2区
文献类型:
--
作者:
Marcellin E;Licona-Cassani C;Mercer TR;Palfreyman RW;Nielsen LK

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准确的细菌基因组注释为理解细胞功能、行为和致病性提供了一个框架,对于代谢工程来说是必不可少的。仅基于计算机预测的注释是不准确的,特别是对于大的、高G + C含量的基因组,因为在基因长度和基因组织方面缺乏相似性来模拟生物体。在这里,我们描述了一种由2D系统生物学驱动的使用蛋白质组学、基因组规模的代谢重构、RNA测序和小RNA测序对Saccharopolyspora Red thraea基因组的重新注释。我们观察了300多个基因间隔区的转录,在基因间隔区检测到59个肽,确认了164个先前被注释为假设蛋白质的开放阅读框,并利用基因组规模的代谢重建将功能重新分配到开放阅读框。最后,我们提出了一种新的方法,通过测序小RNA来定位整个基因组中的核糖体结合位点。这项工作描述了一种新的糖多孢菌基因组注释框架。基于实验观察,2D注释框架大大减少了使用计算预测算法注释大-高G + C含量基因组时经常犯的错误。
Accurate bacterial genome annotations provide a framework to understanding cellular functions, behavior and pathogenicity and are essential for metabolic engineering. Annotations based only on in silico predictions are inaccurate, particularly for large, high G + C content genomes due to the lack of similarities in gene length and gene organization to model organisms. Here we describe a 2D systems biology driven re-annotation of the Saccharopolyspora erythraea genome using proteogenomics, a genome-scale metabolic reconstruction, RNA-sequencing and small-RNA-sequencing. We observed transcription of more than 300 intergenic regions, detected 59 peptides in intergenic regions, confirmed 164 open reading frames previously annotated as hypothetical proteins and reassigned function to open reading frames using the genome-scale metabolic reconstruction. Finally, we present a novel way of mapping ribosomal binding sites across the genome by sequencing small RNAs. The work presented here describes a novel framework for annotation of the Saccharopolyspora erythraea genome. Based on experimental observations, the 2D annotation framework greatly reduces errors that are commonly made when annotating large-high G + C content genomes using computational prediction algorithms.
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