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Single Virus Genomics: A New Tool for Virus Discovery
Whole genome amplification and sequencing of single microbial cells has significantly influenced genomics and microbial ecology by facilitating direct recovery of reference genome data. However, viral genomics continues to suffer due to difficulties related to the isolation and characterization of u...
Autores principales: | , , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3059205/ https://www.ncbi.nlm.nih.gov/pubmed/21436882 http://dx.doi.org/10.1371/journal.pone.0017722 |
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author | Allen, Lisa Zeigler Ishoey, Thomas Novotny, Mark A. McLean, Jeffrey S. Lasken, Roger S. Williamson, Shannon J. |
author_facet | Allen, Lisa Zeigler Ishoey, Thomas Novotny, Mark A. McLean, Jeffrey S. Lasken, Roger S. Williamson, Shannon J. |
author_sort | Allen, Lisa Zeigler |
collection | PubMed |
description | Whole genome amplification and sequencing of single microbial cells has significantly influenced genomics and microbial ecology by facilitating direct recovery of reference genome data. However, viral genomics continues to suffer due to difficulties related to the isolation and characterization of uncultivated viruses. We report here on a new approach called ‘Single Virus Genomics’, which enabled the isolation and complete genome sequencing of the first single virus particle. A mixed assemblage comprised of two known viruses; E. coli bacteriophages lambda and T4, were sorted using flow cytometric methods and subsequently immobilized in an agarose matrix. Genome amplification was then achieved in situ via multiple displacement amplification (MDA). The complete lambda phage genome was recovered with an average depth of coverage of approximately 437X. The isolation and genome sequencing of uncultivated viruses using Single Virus Genomics approaches will enable researchers to address questions about viral diversity, evolution, adaptation and ecology that were previously unattainable. |
format | Text |
id | pubmed-3059205 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-30592052011-03-23 Single Virus Genomics: A New Tool for Virus Discovery Allen, Lisa Zeigler Ishoey, Thomas Novotny, Mark A. McLean, Jeffrey S. Lasken, Roger S. Williamson, Shannon J. PLoS One Research Article Whole genome amplification and sequencing of single microbial cells has significantly influenced genomics and microbial ecology by facilitating direct recovery of reference genome data. However, viral genomics continues to suffer due to difficulties related to the isolation and characterization of uncultivated viruses. We report here on a new approach called ‘Single Virus Genomics’, which enabled the isolation and complete genome sequencing of the first single virus particle. A mixed assemblage comprised of two known viruses; E. coli bacteriophages lambda and T4, were sorted using flow cytometric methods and subsequently immobilized in an agarose matrix. Genome amplification was then achieved in situ via multiple displacement amplification (MDA). The complete lambda phage genome was recovered with an average depth of coverage of approximately 437X. The isolation and genome sequencing of uncultivated viruses using Single Virus Genomics approaches will enable researchers to address questions about viral diversity, evolution, adaptation and ecology that were previously unattainable. Public Library of Science 2011-03-23 /pmc/articles/PMC3059205/ /pubmed/21436882 http://dx.doi.org/10.1371/journal.pone.0017722 Text en Allen et al. http://creativecommons.org/licenses/by/4.0/ 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 properly credited. |
spellingShingle | Research Article Allen, Lisa Zeigler Ishoey, Thomas Novotny, Mark A. McLean, Jeffrey S. Lasken, Roger S. Williamson, Shannon J. Single Virus Genomics: A New Tool for Virus Discovery |
title | Single Virus Genomics: A New Tool for Virus Discovery |
title_full | Single Virus Genomics: A New Tool for Virus Discovery |
title_fullStr | Single Virus Genomics: A New Tool for Virus Discovery |
title_full_unstemmed | Single Virus Genomics: A New Tool for Virus Discovery |
title_short | Single Virus Genomics: A New Tool for Virus Discovery |
title_sort | single virus genomics: a new tool for virus discovery |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3059205/ https://www.ncbi.nlm.nih.gov/pubmed/21436882 http://dx.doi.org/10.1371/journal.pone.0017722 |
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