Reconstructing viral genomes from the environment using fosmid clones: the case of haloviruses

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Título: Reconstructing viral genomes from the environment using fosmid clones: the case of haloviruses
Autor/es: Garcia-Heredia, Inmaculada | Martín Cuadrado, Ana Belén | Mojica, Francisco J.M. | Santos Sánchez, Fernando | Mira, Alex | Anton, Josefa | Rodriguez-Valera, Francisco
Grupo/s de investigación o GITE: Ecología Microbiana Molecular | Microbiología Molecular
Centro, Departamento o Servicio: Universidad de Alicante. Departamento de Fisiología, Genética y Microbiología
Palabras clave: Viral genomes | Environment | Fosmid clones | Haloviruses
Área/s de conocimiento: Microbiología
Fecha de publicación: 30-mar-2012
Editor: Public Library of Science (PLoS)
Cita bibliográfica: GARCIA-HEREDIA, Inmaculada, et al. “Reconstructing viral genomes from the environment using fosmid clones: the case of haloviruses”. PLoS ONE 7(3): e33802. doi:10.1371/journal.pone.0033802
Resumen: Background: Metaviriomes, the viral genomes present in an environment, have been studied by direct sequencing of the viral DNA or by cloning in small insert libraries. The short reads generated by both approaches make it very difficult to assemble and annotate such flexible genomic entities. Many environmental viruses belong to unknown groups or prey on uncultured and little known cellular lineages, and hence might not be present in databases. Methodology and Principal Findings: Here we have used a different approach, the cloning of viral DNA into fosmids before sequencing, to obtain natural contigs that are close to the size of a viral genome. We have studied a relatively low diversity extreme environment: saturated NaCl brines, which simplifies the analysis and interpretation of the data. Forty-two different viral genomes were retrieved, and some of these were almost complete, and could be tentatively identified as head-tail phages (Caudovirales). Conclusions and Significance: We found a cluster of phage genomes that most likely infect Haloquadratum walsbyi, the square archaeon and major component of the community in these hypersaline habitats. The identity of the prey could be confirmed by the presence of CRISPR spacer sequences shared by the virus and one of the available strain genomes. Other viral clusters detected appeared to prey on the Nanohaloarchaea and on the bacterium Salinibacter ruber, covering most of the diversity of microbes found in this type of environment. This approach appears then as a viable alternative to describe metaviriomes in a much more detailed and reliable way than by the more common approaches based on direct sequencing. An example of transfer of a CRISPR cluster including repeats and spacers was accidentally found supporting the dynamic nature and frequent transfer of this peculiar prokaryotic mechanism of cell protection.
Patrocinador/es: This work was supported by projects MAGYK (BIO2008-02444), MICROGEN (Programa CONSOLIDERINGENIO 2010 CDS2009-00006), CGL2'09-12651-C02-01 from the Spanish Ministerio de Ciencia e Innovación, DIMEGEN (PROMETEO/2010/089) and ACOMP/2009/155 from the Generalitat Valenciana. FEDER funds supported this project. IG-H was supported by MAGYK from Ministerio de Ciencia e Innovación. A-BM-C was supported by CONSOLIDER-INGENIO 2010.
ISSN: 1932-6203
DOI: 10.1371/journal.pone.0033802
Idioma: eng
Tipo: info:eu-repo/semantics/article
Derechos: © 2012 Garcia-Heredia et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Revisión científica: si
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Aparece en las colecciones:INV - Microbiología Molecular - Artículos de Revistas
INV - EMM - Artículos de Revistas

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