XB-IMG-116453
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Figure 3. Minimum sequence requirement of S-13 RNA. (A) The sequences used for deletion mutant analysis of S-13 RNA are shown. The mutant names derive from the positions at the start and end of the full-length S-13 RNA. The minimum RNA sequence for binding to mPrrp is indicated at the bottom (positions 26–73). The lower case character sequence at 5′ terminal is derived from the sequence for transcription initiation by T7 RNA polymerase and the other one at the 3′ terminal is derived from the sequence of the restriction enzyme site (BamHI). (B) Gel mobility shift assay of mPrrp-2xRBD protein with deletion mutants. Ten femtomoles of 32P-labeled RNAs (final, 0.5 nM) was incubated with various concentrations of mPrrp-2xRBD protein: 0 nM (lanes 1, 4, 7, 10, 13 and 16), 100 nM (lanes 2, 5, 8, 11, 14 and 17) and 300 nM (lanes 3, 6, 9, 12, 15 and 18), and the mixtures were run on non-denaturing polyacrylamide gels. Free RNA probes and RNA–protein complexes are indicated by arrowheads as in previous figures. 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 Larger Image Printer Friendly View |