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Journal of Virology, September 2000, p. 7803-7813, Vol. 74, No. 17
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

MM1, a Temperate Bacteriophage of the Type 23F Spanish/USA Multiresistant Epidemic Clone of Streptococcus pneumoniae: Structural Analysis of the Site-Specific Integration System

Emmanuel Gindreau,dagger Rubens López,* and Pedro García

Centro de Investigaciones Biológicas, CSIC, Velázquez 144, 28006 Madrid, Spain

Received 23 March 2000/Accepted 23 May 2000

We have characterized a temperate phage (MM1) from a clinical isolate of the multiply antibiotic-resistant Spanish/American 23F Streptococcus pneumoniae clone (Spain23F-1 strain). The 40-kb double-stranded genome of MM1 has been isolated as a DNA-protein complex. The use of MM1 DNA as a probe revealed that the phage genome is integrated in the host chromosome. The host and phage attachment sites, attB and attP, respectively, have been determined. Nucleotide sequencing of the attachment sites identified a 15-bp core site (5'-TTATAATTCATCCGC-3') that has not been found in any bacterial genome described so far. Sequence information revealed the presence of an integrase gene (int), which represents the first identification of an integrase in the pneumococcal system. A 1.5-kb DNA fragment embracing attP and the int gene contained all of the genetic information needed for stable integration of a nonreplicative plasmid into the attB site of a pneumococcal strain. This vector will facilitate the introduction of foreign genes into the pneumococcal chromosome. Interestingly, DNAs highly similar to that of MM1 have been detected in several clinical pneumococcal isolates of different capsular types, suggesting a widespread distribution of these phages in relevant pathogenic strains.

* Corresponding author. Mailing address: Departamento de Microbiología Molecular, Centro de Investigaciones Biológicas, CSIC, Velázquez 144, 28006 Madrid, Spain. Phone: (34-91) 5611800. Fax: (34-91) 5627518. E-mail: ruben{at}cib.csic.es.

dagger Present address: Laboratoire de Biotechnologie et de Microbiologie Appliquée, Faculté d'Oenologie, 33405 Talence Cedex, France.

Journal of Virology, September 2000, p. 7803-7813, Vol. 74, No. 17
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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