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. 1998 Jul 21;95(15):8485–8490. doi: 10.1073/pnas.95.15.8485

Figure 4.

Figure 4

Reticulocyte lysate expression of p133 and telomerase RNA. (A) Telomerase activity was assayed for p133 coexpressed with RNA [p133+RNA] (lane 1); individual components expressed alone [p133], [RNA] (lanes 2 and 3); or p133 with motif C substitution of both aspartic acids for alanines coexpressed with RNA [DDp133+RNA] (lane 4). Assays also were performed for lysate-expressed p133 and RNA mixed after synthesis (lane 5); lysate-expressed p133 mixed with purified telomerase RNA (lanes 6–8: 20 ng, 200 ng, 2 μg of RNA); purified RNA alone (lane 9: 2 μg RNA); or partially purified endogenous telomerase (NT, lane 10). The radiolabel in recombinant enzyme lanes in the top half of the gel is from lysate labeling of plasmid DNA. (B) SDS/PAGE followed by autoradiography was used to detect recombinant protein synthesized in the lysates indicated. The migration of molecular mass markers is indicated at right. (C) Northern analysis of telomerase RNA was used to detect recombinant RNA synthesized in 2.5 μl of the lysates indicated. Telomerase RNA standards were loaded at left (1.1, 0.6, 0.3 ng RNA).