Archaeal viruses of the sulfolobales: isolation, infection, and CRISPR spacer acquisition

Susanne Erdmann; Roger Antony Garrett

doi:10.1007/978-1-4939-2687-9_14

Archaeal viruses of the sulfolobales: isolation, infection, and CRISPR spacer acquisition

Susanne Erdmann, Roger Antony Garrett

Functional Genomics

2 Citations (Scopus)

Abstract

Infection of archaea with phylogenetically diverse single viruses, performed in different laboratories, has failed to activate spacer acquisition into host CRISPR loci. The first successful uptake of archaeal de novo spacers was observed on infection of Sulfolobus solfataricus P2 with an environmental virus mixture isolated from Yellowstone National Park (Erdmann and Garrett, Mol Microbiol 85:1044-1056, 2012). Experimental studies of isolated genetic elements from this mixture revealed that SMV1 (S ulfolobus Monocauda Virus 1), a tailed spindle-shaped virus, can induce spacer acquisition in CRISPR loci of Sulfolobus species from a second coinfecting conjugative plasmid or virus (Erdmann and Garrett, Mol Microbiol 85:1044-1056, 2012; Erdmann et al. Mol Microbiol 91:900-917, 2014). Here we describe, firstly, the isolation of archaeal virus mixtures from terrestrial hot springs and the techniques used both to infect laboratory strains with these virus mixtures and to obtain purified virus particles. Secondly, we present the experimental conditions required for activating SMV1-induced spacer acquisition in two different Sulfolobus species.

Original language	English
Title of host publication	CRISPR : methods and protocols
Editors	Magnus Lundgren, Emmanuelle Charpentier, Peter C. Fineran
Number of pages	10
Publisher	Springer
Publication date	2015
Pages	223-232
ISBN (Print)	978-1-4939-2686-2
ISBN (Electronic)	978-1-4939-2687-9
DOIs	https://doi.org/10.1007/978-1-4939-2687-9_14
Publication status	Published - 2015

Series	Methods in Molecular Biology
Volume	1311
ISSN	1064-3745

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Archaeal viruses of the sulfolobales: isolation, infection, and CRISPR spacer acquisition. / Erdmann, Susanne; Garrett, Roger Antony.
CRISPR: methods and protocols. ed. / Magnus Lundgren; Emmanuelle Charpentier; Peter C. Fineran. Springer, 2015. p. 223-232 (Methods in Molecular Biology, Vol. 1311).

Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review

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abstract = "Infection of archaea with phylogenetically diverse single viruses, performed in different laboratories, has failed to activate spacer acquisition into host CRISPR loci. The first successful uptake of archaeal de novo spacers was observed on infection of Sulfolobus solfataricus P2 with an environmental virus mixture isolated from Yellowstone National Park (Erdmann and Garrett, Mol Microbiol 85:1044-1056, 2012). Experimental studies of isolated genetic elements from this mixture revealed that SMV1 (S ulfolobus Monocauda Virus 1), a tailed spindle-shaped virus, can induce spacer acquisition in CRISPR loci of Sulfolobus species from a second coinfecting conjugative plasmid or virus (Erdmann and Garrett, Mol Microbiol 85:1044-1056, 2012; Erdmann et al. Mol Microbiol 91:900-917, 2014). Here we describe, firstly, the isolation of archaeal virus mixtures from terrestrial hot springs and the techniques used both to infect laboratory strains with these virus mixtures and to obtain purified virus particles. Secondly, we present the experimental conditions required for activating SMV1-induced spacer acquisition in two different Sulfolobus species.",

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AB - Infection of archaea with phylogenetically diverse single viruses, performed in different laboratories, has failed to activate spacer acquisition into host CRISPR loci. The first successful uptake of archaeal de novo spacers was observed on infection of Sulfolobus solfataricus P2 with an environmental virus mixture isolated from Yellowstone National Park (Erdmann and Garrett, Mol Microbiol 85:1044-1056, 2012). Experimental studies of isolated genetic elements from this mixture revealed that SMV1 (S ulfolobus Monocauda Virus 1), a tailed spindle-shaped virus, can induce spacer acquisition in CRISPR loci of Sulfolobus species from a second coinfecting conjugative plasmid or virus (Erdmann and Garrett, Mol Microbiol 85:1044-1056, 2012; Erdmann et al. Mol Microbiol 91:900-917, 2014). Here we describe, firstly, the isolation of archaeal virus mixtures from terrestrial hot springs and the techniques used both to infect laboratory strains with these virus mixtures and to obtain purified virus particles. Secondly, we present the experimental conditions required for activating SMV1-induced spacer acquisition in two different Sulfolobus species.

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Archaeal viruses of the sulfolobales: isolation, infection, and CRISPR spacer acquisition

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