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Genetics and Molecular Biology |
CSIRO Molecular Science, Sydney Laboratory, PO Box 184, North Ryde, NSW 1670, Australia1
School of Biological Sciences, Macquarie University Sydney, NSW 2109, Australia2
Author for correspondence: Ruth M. Hall. Tel: +1 612 9490 5162. Fax: +1 612 9490 5005. e-mail: ruth.hall{at}molsci.csiro.au
Integron-encoded integrases recognize two distinct types of recombination site: attI sites, found in integrons, and members of the 59-base element (59-be) family, found in the integron-associated gene cassettes. The class 1 integron integrase, IntI1, catalyses recombination between attI1 and a 59-be, two 59-be, or two attI1 sites, but events involving two attI1 sites are less efficient than the reactions in which a 59-be participates. The full attI1 site is required for high-efficiency recombination with a 59-be site. It is 65 bp in length and includes a simple site, consisting of a pair of inversely oriented IntI1-binding domains, together with two further directly oriented IntI1-binding sites designated strong and weak. However, a smaller region that contains only the simple site is sufficient to support a lower level of recombination with a complete attI1 partner and the features that determine the orientation of attI1 reside within this region. An unusual reaction between the attI1 site and a 59-be appears to be responsible for the loss of the central region of a 59-be to create a potential fusion of two adjacent gene cassettes.
Keywords: attI, integron, integrase, site-specific recombination, gene cassettes
Abbreviations: 2°rs, secondary recombination site; 59-be, 59-base element; 5'-CS, 5' conserved segment; Ap, ampicillin; Cm, chloramphenicol; IntI1, integrase of class 1 integrons; Nx, nalidixic acid; Sm, streptomycin; Su, sulphamethoxazole; Tc, tetracycline; Tp, trimethoprim
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