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Volume 147, Issue 8

Resolvase-like recombination performed by the TP901-1 integrase

The GenBank accession number for the sequence reported in this paper is Y15043.

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Abstract

The site-specific recombination system of temperate lactococcal bacteriophage TP901-1 is unusual in several respects. First, the integrase belongs to the family of extended resolvases rather than to the λ integrase family and second, in the presence of this integrase, a 56 bp fragment is sufficient for efficient recombination with the chromosomal site in the host subsp. MG1363. In the present work, this site was analysed and a 43 bp region was found to be the smallest fragment able to participate fully in recombination. studies showed that the TP901-1 integrase binds this 43 bp fragment, the 56 bp and a larger fragment with equal affinity. Mutational analysis of the 5 bp common core region (TCAAT) showed that the TC dinucleotide is essential for recombination, but not for binding of the integrase, whereas none of the last three bases are important for recombination. When a number of sites, obtained by recombination between an site containing a mutation in this TC dinucleotide and a wild-type site, were sequenced, a mix of sites with the wild-type or the mutated sequence was obtained. These results are consistent with the hypothesis that the TC dinucleotide constitutes the TP901-1 overlap region. A 2 bp overlap region has been observed in recombination reactions catalysed by all other members of the resolvase/invertase family tested so far. By selecting for sites with a decreased ability to participate in recombination, two bases located outside the core region of were shown to be involved in the binding of the TP901-1 integrase.

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Keyword(s): Ap, ampicillin , chromosomal attachment site , Cm, chloramphenicol , Ery, erythromycin , extended resolvase , Kn, kanamycin , lactococcal bacteriophage , mechanism of recombination and site-specific recombination
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2001-08-01
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Resolvase-like recombination performed by the TP901-1 integraseThe GenBank accession number for the sequence reported in this paper is Y15043.
Microbiology 147, 2051 (2001); https://doi.org/10.1099/00221287-147-8-2051
/content/journal/micro/10.1099/00221287-147-8-2051
/content/journal/micro/10.1099/00221287-147-8-2051
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