The 3′ end of ISY becomes covalently linked to the target DNA in the strand transfer reaction

<p><b>Copyright information:</b></p><p>Taken from " transposition of ISY, a bacterial insertion sequence belonging to the Tc/ family"</p><p></p><p>Molecular Microbiology 2007;65(6):1432-1443.</p><p>Published online Jan 2007</p><p>PMCID:PMC2170065.</p><p>© 2007 The Authors; Journal compilation © 2007 Blackwell Publishing Ltd</p> A. A pre-cleaved transposon end, 5′ end-labelled on either the top or bottom strand, was incubated with transposase and a supercoiled target plasmid. Samples were withdrawn at the indicated times and run on a non-denaturing agarose gel (TAE) or a strand-separating alkaline gel (NaOH). DNA was detected by ethidium staining, and by autoradiography (P). The positions of supercoiled (sc) and open circle (oc) target plasmid are shown to the left of the ethidium-stained gel. The positions of single end-joined (SEJ) and double end-joined (DEJ) strand transfer products are shown to the right of the autoradiographs. SEJ and DEJ products are indistinguishable on the strand separating gel. Unreacted labelled substrate has been cropped from the bottom of both autoradiographs. B. Sequence of the pre-cleaved end and schematic diagram of the reaction products

Binding of N-terminal transposase derivatives to the ISY inverted repeat

<p><b>Copyright information:</b></p><p>Taken from " transposition of ISY, a bacterial insertion sequence belonging to the Tc/ family"</p><p></p><p>Molecular Microbiology 2007;65(6):1432-1443.</p><p>Published online Jan 2007</p><p>PMCID:PMC2170065.</p><p>© 2007 The Authors; Journal compilation © 2007 Blackwell Publishing Ltd</p> A. Transposase deletion derivatives were purified by metal affinity chromatography and analysed by Coomassie-stained Tris-tricine SDS-PAGE. B. The indicated protein (1.25 μM) was incubated with IRL58, and protein–DNA complexes were separated by non-denaturing polyacrylamide gel electrophoresis. C. The indicated concentrations of Tnp and Tnp were incubated with IRL58 and complexes were separated as in (B)