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. 2009 Apr 30;37(12):3981–3989. doi: 10.1093/nar/gkp271

Figure 2.

Figure 2.

Activity assays of ribozymes lacking a native IGS. (A) Splicing activity of the Azoarcus L–30 construct V•X•Y•Z when supplied with an exogenous substrate that either contains the IGS sequence GUG (h•V, h•Y and h•Z) or that does not (h•X and SNL-1a). Upon splicing, the 5′-radiolabeled ribozyme is expected to append the 3′ portion of the substrate to its own 3′ end, producing a product of the sizes indicated by the small arrows; no splicing is visible with h•X or SNL-1a, which should give products of lower molecular weight than the product with h•V. (B) Bipartate covalent self-assembly reactions. In each set, the V-containing fragment is 5′ radiolabeled. The R2F2 reactions were designed with splice junctions (V–X or Y–Z) that can only form a covalent bond via a two-step recombination reaction, while the tF2 reactions were designed with splice junctions that can form a covalent bond through either a two-step recombination or through a one-step recombination (22). This accounts for the slight size difference between the reactants or products in each system. The covalent self-assembly product V•X•Y•Z (∼175 nt) is indicated.