Isolation, growth and genome of the Rhodothermus RM378 thermophilic bacteriophage
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Several bacteriophages that infect different strains of the thermophilic bacterium Rhodothermus marinus were isolated and their infection pattern was studied. One phage, named RM378 was cultivated and characterized. The RM378 genome was also sequenced and analyzed. The phage was grouped as a member of the Myoviridae family with A2 morphology. It had a moderately elongated head, with dimensions of 85 and 95 nm between opposite apices and a 150 nm long tail, attached with a connector to the head. RM378 showed a virulent behavior that followed a lytic cycle of infection. It routinely gave lysates with 1011 pfu/ml, and sometimes reached titers as high as 1013 pfu/ml. The titer remained stable up to 65 °C but the phage lost viability when incubated at higher temperatures. Heating for 30 min at 96 °C lowered the titer by 104. The RM378 genome consisted of ds DNA of 129.908 bp with a GC ratio of 42.0 % and contained about 120 ORFs. A few structural proteins, such as the major head protein corresponding to the gp23 in T4, could be identified. Only 29 gene products as probable homologs to other proteins of known function could be predicted, with most showing only low similarity to known proteins in other bacteriophages. These and other studies based on sequence analysis of a large number of phage genomes showed RM378 to be distantly related to all other known T4-like phages.
KeywordsThermophile Bacteriophage RM378 Rhodothermus marinus
Plaque forming units
Parts of the research leading to these results have received funding from the European Union’s Seventh Framework Programme managed by REAResearch Executive Agency http://ec.europa.eu/research/rea [(FP7/2007-2013) (FP7/2007-2011)] under grant agreement no 286556. We thank Johann Arnfinnsson and Gudmundur Georgsson for the electron microscopy, and Haflidi Asgrimsson and Brynjolfur Bjarnason for software development.
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