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Captured metagenomics: large-scale targeting of genes based on ‘sequence capture’ reveals functional diversity in soils

Microbial enzyme diversity is a key to understand many ecosystem processes. Whole metagenome sequencing (WMG) obtains information on functional genes, but it is costly and inefficient due to large amount of sequencing that is required. In this study, we have applied a captured metagenomics technique...

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Detalles Bibliográficos
Autores principales: Manoharan, Lokeshwaran, Kushwaha, Sandeep K., Hedlund, Katarina, Ahrén, Dag
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4675713/
https://www.ncbi.nlm.nih.gov/pubmed/26490729
http://dx.doi.org/10.1093/dnares/dsv026
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author Manoharan, Lokeshwaran
Kushwaha, Sandeep K.
Hedlund, Katarina
Ahrén, Dag
author_facet Manoharan, Lokeshwaran
Kushwaha, Sandeep K.
Hedlund, Katarina
Ahrén, Dag
author_sort Manoharan, Lokeshwaran
collection PubMed
description Microbial enzyme diversity is a key to understand many ecosystem processes. Whole metagenome sequencing (WMG) obtains information on functional genes, but it is costly and inefficient due to large amount of sequencing that is required. In this study, we have applied a captured metagenomics technique for functional genes in soil microorganisms, as an alternative to WMG. Large-scale targeting of functional genes, coding for enzymes related to organic matter degradation, was applied to two agricultural soil communities through captured metagenomics. Captured metagenomics uses custom-designed, hybridization-based oligonucleotide probes that enrich functional genes of interest in metagenomic libraries where only probe-bound DNA fragments are sequenced. The captured metagenomes were highly enriched with targeted genes while maintaining their target diversity and their taxonomic distribution correlated well with the traditional ribosomal sequencing. The captured metagenomes were highly enriched with genes related to organic matter degradation; at least five times more than similar, publicly available soil WMG projects. This target enrichment technique also preserves the functional representation of the soils, thereby facilitating comparative metagenomics projects. Here, we present the first study that applies the captured metagenomics approach in large scale, and this novel method allows deep investigations of central ecosystem processes by studying functional gene abundances.
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spelling pubmed-46757132015-12-11 Captured metagenomics: large-scale targeting of genes based on ‘sequence capture’ reveals functional diversity in soils Manoharan, Lokeshwaran Kushwaha, Sandeep K. Hedlund, Katarina Ahrén, Dag DNA Res Full Papers Microbial enzyme diversity is a key to understand many ecosystem processes. Whole metagenome sequencing (WMG) obtains information on functional genes, but it is costly and inefficient due to large amount of sequencing that is required. In this study, we have applied a captured metagenomics technique for functional genes in soil microorganisms, as an alternative to WMG. Large-scale targeting of functional genes, coding for enzymes related to organic matter degradation, was applied to two agricultural soil communities through captured metagenomics. Captured metagenomics uses custom-designed, hybridization-based oligonucleotide probes that enrich functional genes of interest in metagenomic libraries where only probe-bound DNA fragments are sequenced. The captured metagenomes were highly enriched with targeted genes while maintaining their target diversity and their taxonomic distribution correlated well with the traditional ribosomal sequencing. The captured metagenomes were highly enriched with genes related to organic matter degradation; at least five times more than similar, publicly available soil WMG projects. This target enrichment technique also preserves the functional representation of the soils, thereby facilitating comparative metagenomics projects. Here, we present the first study that applies the captured metagenomics approach in large scale, and this novel method allows deep investigations of central ecosystem processes by studying functional gene abundances. Oxford University Press 2015-12 2015-10-21 /pmc/articles/PMC4675713/ /pubmed/26490729 http://dx.doi.org/10.1093/dnares/dsv026 Text en © The Author 2015. Published by Oxford University Press on behalf of Kazusa DNA Research Institute. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Full Papers
Manoharan, Lokeshwaran
Kushwaha, Sandeep K.
Hedlund, Katarina
Ahrén, Dag
Captured metagenomics: large-scale targeting of genes based on ‘sequence capture’ reveals functional diversity in soils
title Captured metagenomics: large-scale targeting of genes based on ‘sequence capture’ reveals functional diversity in soils
title_full Captured metagenomics: large-scale targeting of genes based on ‘sequence capture’ reveals functional diversity in soils
title_fullStr Captured metagenomics: large-scale targeting of genes based on ‘sequence capture’ reveals functional diversity in soils
title_full_unstemmed Captured metagenomics: large-scale targeting of genes based on ‘sequence capture’ reveals functional diversity in soils
title_short Captured metagenomics: large-scale targeting of genes based on ‘sequence capture’ reveals functional diversity in soils
title_sort captured metagenomics: large-scale targeting of genes based on ‘sequence capture’ reveals functional diversity in soils
topic Full Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4675713/
https://www.ncbi.nlm.nih.gov/pubmed/26490729
http://dx.doi.org/10.1093/dnares/dsv026
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