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Applied and Environmental Microbiology, December 2008, p. 7552-7560, Vol. 74, No. 24
0099-2240/08/$08.00+0     doi:10.1128/AEM.01709-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Isolation of New Stenotrophomonas Bacteriophages and Genomic Characterization of Temperate Phage S1{triangledown}

Pilar García,1,2 Cristina Monjardín,1 Rebeca Martín,1 Carmen Madera,1 Nora Soberón,1 Eva Garcia,3 Álvaro Meana,3 and Juan E. Suárez1,2*

Área de Microbiología, Facultad de Medicina, Universidad de Oviedo, Oviedo, Spain,1 Instituto de Productos Lácteos de Asturias (CSIC), Villaviciosa, Spain,2 Centro Comunitario de Sangre y Tejidos and CIBER de Enfermedades Raras (CIBERER), Oviedo, Spain3

Received 24 July 2008/ Accepted 15 October 2008

Twenty-two phages that infect Stenotrophomonas species were isolated through sewage enrichment and prophage induction. Of them, S1, S3, and S4 were selected due to their wide host ranges compared to those of the other phages. S1 and S4 are temperate siphoviruses, while S3 is a virulent myovirus. The genomes of S3 and S4, about 33 and 200 kb, were resistant to restriction digestion. The lytic cycles lasted 30 min for S3 and about 75 min for S1 and S4. The burst size for S3 was 100 virions/cell, while S1 and S4 produced about 75 virus particles/cell. The frequency of bacteriophage-insensitive host mutants, calculated by dividing the number of surviving colonies by the bacterial titer of a parallel, uninfected culture, ranged between 10–5 and 10–6 for S3 and 10–3 and 10–4 for S1 and S4. The 40,287-bp genome of S1 contains 48 open reading frames (ORFs) and 12-bp 5' protruding cohesive ends. By using a combination of bioinformatics and experimental evidence, functions were ascribed to 21 ORFs. The morphogenetic and lysis modules are well-conserved, but no lysis-lysogeny switch or DNA replication gene clusters were recognized. Two major clusters of genes with respect to transcriptional orientation were observed. Interspersed among them were lysogenic conversion genes encoding phosphoadenosine phosphosulfate reductase and GspM, a protein involved in the general secretion system II. The attP site of S1 may be located within a gene that presents over 75% homology to a Stenotrophomonas chromosomal determinant.

* Corresponding author. Mailing address: Área de Microbiología, Facultad de Medicina, Universidad de Oviedo, Asturias, Spain. Phone: 34 985 103559. Fax: 34 985 103148. E-mail: evaristo{at}uniovi.es

{triangledown} Published ahead of print on 24 October 2008.

Applied and Environmental Microbiology, December 2008, p. 7552-7560, Vol. 74, No. 24
0099-2240/08/$08.00+0     doi:10.1128/AEM.01709-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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