Single Virus Genomics: A New Tool for Virus Discovery

• Published in: PloS one Vol. 6; no. 3; p. e17722
• Main Authors: Allen, Lisa Zeigler, Ishoey, Thomas, Novotny, Mark A., McLean, Jeffrey S., Lasken, Roger S., Williamson, Shannon J.
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 23.03.2011; Public Library of Science (PLoS)
• Subjects: Adaptations; Algorithms; Amplification; Bacteria; Bacteriophage lambda - genetics; Bacteriophage lambda - isolation & purification; Bacteriophage T4 - genetics; Bacteriophage T4 - isolation & purification; Biodiversity; Biological evolution; Biology; Cloning; Data processing; Data recovery; Deoxyribonucleic acid; DNA; DNA sequencing; E coli; Ecological effects; Ecology; Escherichia coli; Evolution; Flow Cytometry; Gene sequencing; Genetic aspects; Genetic Loci - genetics; Genome, Viral - genetics; Genomes; Genomics; Genomics - methods; Microorganisms; Microscopy; Microscopy, Confocal; Phages; Polymerase Chain Reaction; Principal components analysis; Reference Standards; Reproducibility of Results; Sequence Analysis, DNA; Venter, J Craig; Viruses; United States > US; California
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0017722

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.