A Two-Piece Derivative of a Group I Intron RNA as a Platform for Designing Self-Assembling RNA Templates to Promote Peptide Ligation
Figure 5
Dissection of the template RNA complex (P5-λboxB : Ez-RRE) for the chemical ligation between λN-peptide1 and rev-peptide2. (a) Time course of ligation reaction (with 7.5 μM each peptide) in the presence of 10 mM Mg2+. Reactions were carried out with template RNA having PRM (P5-λboxB : Ez-RRE) and lacking PRM (P5 : Ez), and also without RNA template. (b) The contribution of the template subunit having PRM (P5-λboxB and Ez-RRE subunits). Reactions were carried out for 7 h in the presence of 10 mM Mg2+ ions. (c) Effects of PRMs on the P5 : Ez complex formation. Mobility shift assay was carried out with native gal electrophoresis in the presence of 10 mM Mg2+ ions. To visualize RNA molecules on the gel, the 3′ ends of FL-labeled RNAs were labeled with BODIPY. (d) The contribution of PRMs in the template ability of the two-piece RNA complex (P5-λboxB : Ez-RRE). Reactions were carried out for 7 h in the presence of 10 mM Mg2+ ions.
