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Two of these colonies, G4ST-2-41 and G4ST-2-42, were transferred to liquid culture, and after 2 days, cells from 1 ml of each were harvested by centrifugation (5 min at 6,000 × ), washed three times in nutrient-free medium, and then plated.Twelve colonies each of G4ST-2-41 and G4ST-2-42 were picked, restreaked, and then shown by PCR to continuously carry STIV2.Samples were taken from the hot spring Ice G4 (88.3°C, p H 3.5) in the Hverakjalki valley, Hveragerdi, Iceland, in April 2006.A 1-ml sample of both liquid and silt was diluted in 50 ml of ST medium and shaken for 1 week at 78°C with aeration.A 1-ml aliquot was transferred to 50 ml of fresh ST medium and incubated for a further 4 days. One round of colony purification was carried out, and 50 colonies were grown in 5 ml of ST medium.STIV2 was observed in the supernatant of strain G4ST-2 in transmission electron microscopy (TEM) by negative staining of the sample (Fig. In order to identify the host, its 16S r RNA gene was amplified by PCR and sequenced (primers 8a F, TCYGGTTGATCCTGCC, and 1512u R, ACGGHTACCTTGTTACGACTT). Shown is a cryo-electron micrograph taken at 2.9-μm underfocus showing STIV2 viral particles (black arrow).These viruses, of the so-called “PRD1-adeno lineage,” are suggested to have a common ancestor that precedes the division of the three domains of life (5).Here we report the isolation, genome sequence, identification of the major virion proteins, and three-dimensional structural characterization of a novel, icosahedrally symmetric archaeal virus.

Colony PCR was used to detect the presence of STIV2 in all of the colonies.

Characterization of this new isolate revealed it to be similar to STIV on the levels of genome and structural organization.

The genome organization indicates that these two viruses have diverged from a common ancestor.

Known archaeal viruses (∼40) constitute less than 1% of the ∼8,000 species of viruses that have been described (13, 59).

So far, virtually all the archaeal viruses have double-stranded DNA (ds DNA) genomes, with the first viral single-stranded DNA (ss DNA) genome being the most recently described (46, 61).

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We demonstrate that, despite overall architectural similarity to STIV, the turreted host attachment structures of these two viruses differ significantly.

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