Characterization of the attP site of the integrative element pSAM2 from Streptomyces ambofaciens.
pSAM2 is integrated into the Streptomyces ambofaciens chromosome through site-specific recombination between the element (attP) and the chromosomal (attB) site. The 43 kDa integrase protein encoded by pSAM2 catalyses this recombination event. Tools have been developed to study site-specific recombination in Escherichia coli. In vivo studies showed that a 360 bp fragment of attP is required for efficient site-specific recombination and that int can be provided in trans. pSAM2 integrase was purified and overexpressed in E. coli and Int binding at the attP site was studied. DNaseI footprinting revealed two sites that bind integrase strongly and appear to be symmetrical with regard to the core site. These two P1/P2 arm-type sites both contain a 17 bp motif that is identical except at one position, GTCACGCAG(A/T)TAGACAC. P1 and P2 are essential for site-specific recombination.
Raynal
Alain
A
Laboratoire de Biologie et Génétique Moléculaire, Institut de Génétique et Microbiologie, UMR CNRS 8621, Bât. 400, Université Paris-Sud, F-91405 Orsay Cedex, France1.
Friedmann
Annick
A
Laboratoire de Biologie et Génétique Moléculaire, Institut de Génétique et Microbiologie, UMR CNRS 8621, Bât. 400, Université Paris-Sud, F-91405 Orsay Cedex, France1.
Tuphile
Karine
K
Laboratoire de Biologie et Génétique Moléculaire, Institut de Génétique et Microbiologie, UMR CNRS 8621, Bât. 400, Université Paris-Sud, F-91405 Orsay Cedex, France1.
Guerineau
Michel
M
Laboratoire de Biologie et Génétique Moléculaire, Institut de Génétique et Microbiologie, UMR CNRS 8621, Bât. 400, Université Paris-Sud, F-91405 Orsay Cedex, France1.
Pernodet
Jean-Luc
JL
Laboratoire de Biologie et Génétique Moléculaire, Institut de Génétique et Microbiologie, UMR CNRS 8621, Bât. 400, Université Paris-Sud, F-91405 Orsay Cedex, France1.
D001287:Attachment Sites, Microbiological; D001483:Base Sequence; D001665:Binding Sites; D018983:DNA Footprinting; D004251:DNA Transposable Elements / Q000235:genetics* / Q000502:physiology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D011995:Recombination, Genetic; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism
D004251:DNA Transposable Elements; D019426:Integrases