Terbium(III)-mediated footprinting of the four 3′-P-labeled -acting genomic HDV ribozyme variants

Abstract

<p><b>Copyright information:</b></p><p>Taken from "The genomic HDV ribozyme utilizes a previously unnoticed U-turn motif to accomplish fast site-specific catalysis"</p><p></p><p>Nucleic Acids Research 2007;35(6):1933-1946.</p><p>Published online 2 Mar 2007</p><p>PMCID:PMC1874588.</p><p>© 2007 The Author(s)</p> () Control experiment of background cleavage in the presence of Tb. The HDV ribozyme N − 1 variants were incubated for 30 min at 22°C under the indicated ionic conditions (see also Materials and methods section). We then separated the 88-nt reaction precursor from the 85-nt, faster migrating 3′-product, as identified by comparison with purified 3′-product from self-cleavage of the U − 1, A − 1 and G − 1 variants (3′P-N − 1). ‘Fresh’ indicates that the material was not incubated before loading onto the gel. () Terbium(III)-mediated footprinting of genomic HDV ribozyme variants. As in (A), the HDV ribozyme N − 1 variants were incubated for 30 min at 22°C under the indicated ionic conditions and then analyzed on a sequencing gel alongside alkaline hydrolysis (OH) and G-specific RNase T1 ladders for sequence identification (see also Materials and methods section). ‘Fresh’ indicates that the material was not incubated before loading onto the gel