Figure 2. Binding experiments on several selected RNAs and mPrrp-2xRBD protein. (A) Gel mobility shift assay of mPrrp-2xRBD protein with several selected RNAs (S-2, lanes 1–4; S-13, lanes 5–8; and S-52, lanes 9–12). An aliquot of 10 fmol of 32P-labeled RNAs (final, 0.5 nM) was incubated with various concentrations of mPrrp-2xRBD protein: 0 nM (lanes 1, 5 and 9), 50 nM (lanes 2, 6 and 10), 100 nM (lanes 3, 7 and 11) and 300 nM (lanes 4, 8 and 12), and the mixtures were run on non-denaturing polyacrylamide gels. (B) Competitive RNA-binding experiments involving unlabeled RNAs. Ten femtomoles (final, 0.5 nM) of 32P-labeled S-13 RNA was incubated with mPrrp-2xRBD protein at a final concentration of 300 nM, and the following amounts of unlabeled competitor RNAs: 100 fmol (lanes 3 and 6), 1000 fmol (lanes 4 and 7) and 10 000 fmol (lanes 5 and 8). Free RNA probes and RNA–protein complexes are indicated by arrowheads. pBS RNA was transcribed from pBluescript SK(+) digested with XbaI.
Image published in: Hori T et al. (2005)
Image downloaded from an Open Access article in PubMed Central. © 2005, the authors Nucleic Acids Research, Vol. 33 No. 1 © Oxford University Press 2005; all rights reserved
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