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Broad-Host-Range Yersinia Phage PY100: Genome Sequence, Proteome Analysis of Virions, and DNA Packaging Strategy

Dominik Schwudke,² Asgar Ergin,^3, Kathrin Michael,^3, Sven Volkmar,³ Bernd Appel,¹ Dorothea Knabner,¹ Antje Konietzny,¹ and Eckhard Strauch^1*

Bundesinstitut für Risikobewertung, Federal Institute for Risk Assessment, Diedersdorfer Weg 1, 12277 Berlin, Germany,¹ Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstr. 108, 01307 Dresden, Germany,² Robert Koch Institute, Nordufer 20, 13353 Berlin, Germany³

Received 29 August 2007/ Accepted 12 October 2007

PY100 is a lytic bacteriophage with a broad host range within the genus Yersinia. The phage forms plaques on strains of the three human pathogenic species Yersinia enterocolitica, Y. pseudotuberculosis, and Y. pestis at 37°C. PY100 was isolated from farm manure and intended to be used in phage therapy trials. PY100 has an icosahedral capsid containing double-stranded DNA and a contractile tail. The genome consists of 50,291 bp and is predicted to contain 93 open reading frames (ORFs). PY100 gene products were found to be homologous to the capsid proteins and proteins involved in DNA metabolism of the enterobacterial phage T1; PY100 tail proteins possess homologies to putative tail proteins of phage Aa23 of Actinobacillus actinomycetemcomitans. In a proteome analysis of virion particles, 15 proteins of the head and tail structures were identified by mass spectrometry. The putative gene product of ORF2 of PY100 shows significant homology to the gene 3 product (small terminase subunit) of Salmonella phage P22 that is involved in packaging of the concatemeric phage DNA. The packaging mechanism of PY100 was analyzed by hybridization and sequence analysis of DNA isolated from virion particles. Newly replicated PY100 DNA is cut initially at a pac recognition site, which is located in the coding region of ORF2.

Published ahead of print on 26 October 2007.

Present address: Universitätsmedizin Berlin, Charité, Campus Benjamin Franklin, Hindenburgdamm 30, 12200 Berlin, Germany.

Present address: Friedrich-Loeffler-Institut, Südufer 10, 17493 Greifswald-Insel Riems, Germany.