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GENETICS AND MOLECULAR BIOLOGY

Spontaneous Excision of the Salmonella enterica Serovar Enteritidis-Specific Defective Prophage-Like Element φSE14

Carlos A. Santiviago, Carlos J. Blondel, Carolina P. Quezada, Cecilia A. Silva, Pia M. Tobar, Steffen Porwollik, Michael McClelland, Helene L. Andrews-Polymenis, Cecilia S. Toro, Mercedes Zaldívar, Inés Contreras
Carlos A. Santiviago
1Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias Químicas y Farmacéuticas, Universidad de Chile, Santiago, Chile
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  • For correspondence: csantiviago@gmail.com
Carlos J. Blondel
1Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias Químicas y Farmacéuticas, Universidad de Chile, Santiago, Chile
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Carolina P. Quezada
1Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias Químicas y Farmacéuticas, Universidad de Chile, Santiago, Chile
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Cecilia A. Silva
1Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias Químicas y Farmacéuticas, Universidad de Chile, Santiago, Chile
2Programa de Microbiología y Micología, Instituto de Ciencias Biomédicas, Facultad de Medicina, Universidad de Chile, Santiago, Chile
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Pia M. Tobar
1Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias Químicas y Farmacéuticas, Universidad de Chile, Santiago, Chile
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Steffen Porwollik
3Vaccine Research Institute of San Diego, San Diego, California 92121
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Michael McClelland
3Vaccine Research Institute of San Diego, San Diego, California 92121
4Department of Pathology and Laboratory Medicine, College of Health Sciences, University of California, Irvine, Irvine, California 92697
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Helene L. Andrews-Polymenis
5Department of Microbial and Molecular Pathogenesis, College of Medicine, Texas A&M University System Health Science Center, 407 Reynolds Medical Building, College Station, Texas 77843
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Cecilia S. Toro
2Programa de Microbiología y Micología, Instituto de Ciencias Biomédicas, Facultad de Medicina, Universidad de Chile, Santiago, Chile
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Mercedes Zaldívar
1Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias Químicas y Farmacéuticas, Universidad de Chile, Santiago, Chile
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Inés Contreras
1Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias Químicas y Farmacéuticas, Universidad de Chile, Santiago, Chile
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DOI: 10.1128/JB.00270-09
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ABSTRACT

Salmonella enterica serovar Enteritidis has emerged as a major health problem worldwide in the last few decades. DNA loci unique to S. Enteritidis can provide markers for detection of this pathogen and may reveal pathogenic mechanisms restricted to this serovar. An in silico comparison of 16 Salmonella genomic sequences revealed the presence of an ∼12.5-kb genomic island (GEI) specific to the sequenced S. Enteritidis strain NCTC13349. The GEI is inserted at the 5′ end of gene ydaO (SEN1377), is flanked by 308-bp imperfect direct repeats (attL and attR), and includes 21 open reading frames (SEN1378 to SEN1398), encoding primarily phage-related proteins. Accordingly, this GEI has been annotated as the defective prophage SE14 in the genome of strain NCTC13349. The genetic structure and location of φSE14 are conserved in 99 of 103 wild-type strains of S. Enteritidis studied here, including reference strains NCTC13349 and LK5. Notably, an extrachromosomal circular form of φSE14 was detected in every strain carrying this island. The presence of attP sites in the circular forms detected in NCTC13349 and LK5 was confirmed. In addition, we observed spontaneous loss of a tetRA-tagged version of φSE14, leaving an empty attB site in the genome of strain NCTC13349. Collectively, these results demonstrate that φSE14 is an unstable genetic element that undergoes spontaneous excision under standard growth conditions. An internal fragment of φSE14 designated Sdf I has been used as a serovar-specific genetic marker in PCR-based detection systems and as a tool to determine S. Enteritidis levels in experimental infections. The instability of this region may require a reassessment of its suitability for such applications.

  • Copyright © 2010 American Society for Microbiology
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Spontaneous Excision of the Salmonella enterica Serovar Enteritidis-Specific Defective Prophage-Like Element φSE14
Carlos A. Santiviago, Carlos J. Blondel, Carolina P. Quezada, Cecilia A. Silva, Pia M. Tobar, Steffen Porwollik, Michael McClelland, Helene L. Andrews-Polymenis, Cecilia S. Toro, Mercedes Zaldívar, Inés Contreras
Journal of Bacteriology Mar 2010, 192 (8) 2246-2254; DOI: 10.1128/JB.00270-09

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Spontaneous Excision of the Salmonella enterica Serovar Enteritidis-Specific Defective Prophage-Like Element φSE14
Carlos A. Santiviago, Carlos J. Blondel, Carolina P. Quezada, Cecilia A. Silva, Pia M. Tobar, Steffen Porwollik, Michael McClelland, Helene L. Andrews-Polymenis, Cecilia S. Toro, Mercedes Zaldívar, Inés Contreras
Journal of Bacteriology Mar 2010, 192 (8) 2246-2254; DOI: 10.1128/JB.00270-09
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KEYWORDS

prophages
Salmonella enteritidis

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