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Journal of Bacteriology, January 2002, p. 177-182, Vol. 184, No. 1
0021-9193/01/$04.00+0     DOI: 10.1128/JB.184.1.177-182.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Site-Specific Integrative Elements of Rhizobiophage 16-3 Can Integrate into Proline tRNA (CGG) Genes in Different Bacterial Genera

Szabolcs Semsey,^1, Béla Blaha,² Krisztián Köles,¹ László Orosz,^1,2 and Péter P. Papp^1,2*

Institute of Genetics, Agricultural Biotechnology Center, Gödöllö, Szent-Györgyi A. 4., H-2100,¹ Department of Genetics, Faculty of Science, Lóránd Eötvös University, Budapest, Pázmány P. 1/C., H-1117, Hungary²

Received 27 June 2001/ Accepted 27 September 2001

The integrase protein of the Rhizobium meliloti 41 phage 16-3 has been classified as a member of the Int family of tyrosine recombinases. The site-specific recombination system of the phage belongs to the group in which the target site of integration (attB) is within a tRNA gene. Since tRNA genes are conserved, we expected that the target sequence of the site-specific recombination system of the 16-3 phage could occur in other species and integration could take place if the required putative host factors were also provided by the targeted cells. Here we report that a plasmid (pSEM167) carrying the attP element and the integrase gene (int) of the phage can integrate into the chromosomes of R. meliloti 1021 and eight other species. In all cases integration occurred at so-far-unidentified, putative proline tRNA (CGG) genes, indicating the possibility of their common origin. Multiple alignment of the sequences suggested that the location of the att core was different from that expected previously. The minimal attB was identified as a 23-bp sequence corresponding to the anticodon arm of the tRNA.

Present address: Laboratory of Molecular Biology, National Cancer Institute, NIH, Bethesda, MD 20892-4255.

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