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Journal of Bacteriology, November 1999, p. 7034-7042, Vol. 181, No. 22
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

TPW22, a Lactococcal Temperate Phage with a Site-Specific Integrase Closely Related to Streptococcus thermophilus Phage Integrases

Anne Petersen,1 Jytte Josephsen,1,* and Mads G. Johnsen2

Department of Dairy and Food Science, Food Microbiology, The Royal Veterinary and Agricultural University, DK-1958 Frederiksberg C,1 and The Biotechnological Institute, DK-2970 Hørsholm,2 Denmark

Received 21 July 1998/Accepted 22 August 1999

The temperate phage TPW22, induced from Lactococcus lactis subsp. cremoris W22, and the evolutionarily interesting integrase of this phage were characterized. Phage TPW22 was propagated lytically on L. lactis subsp. cremoris 3107, which could also be lysogenized by site-specific integration. The attachment site (attP), 5'-TAAGGCGACGGTCG-3', of phage TPW22 was present on a 7.5-kb EcoRI fragment, a 3.4-kb EcoRI-HindIII fragment of which was sequenced. Sequence information revealed the presence of an integrase gene (int). The deduced amino acid sequence showed 42 and 28% identity with integrases of streptococcal and lactococcal phages, respectively. The identities with these integrase-encoding genes were 52 and 45%, respectively, at the nucleotide level. This could indicate horizontal gene transfer. A stable integration vector containing attP and int was constructed, and integration in L. lactis subsp. cremoris MG1363 was obtained. The existence of an exchangeable lactococcal phage integration module was suggested. The proposed module covers the phage attachment site, the integrase gene, and surrounding factor-independent terminator structures. The phages phi LC3, TP901-1, and TPW22 all have different versions of this module. Phylogenetically, the TPW22 Int links the phi LC3 lactococcal integrase with known Streptococcus thermophilus integrases.

* Corresponding author. Mailing address: Department of Dairy and Food Science, Food Microbiology, The Royal Veterinary and Agricultural University, Rolighedsvej 30, DK-1958 Frederiksberg C, Denmark. Phone: 45 35 28 32 32. Fax: 45 35 28 32 14. E-mail: jjoseph{at}mli.kvl.dk.

Journal of Bacteriology, November 1999, p. 7034-7042, Vol. 181, No. 22
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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