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. 2001 May 1;29(9):1915–1925. doi: 10.1093/nar/29.9.1915

Figure 6.

Figure 6

Binding of the RNase III dsRBD to R1.1[WC] RNA is resistant to EB. A gel shift assay was performed using 5′-32P-labeled R1.1[WC] RNA and purified dsRBD, as described in Materials and Methods. We have shown elsewhere that the RNase III dsRBD binds R1.1[WC] RNA with a Kd of ∼800 nM (A.K.Amarasinghe, S.Su, W.Sun, R.W.Simons and A.W.Nicholson, manuscript in preparation). CaCl2 (10 mM) was included in the binding reaction and gel and running buffers. The reaction was analyzed by phosphorimaging. The dsRBD concentration was 800 nM. The positions of bound and free R1.1[WC] RNA are indicated. The smear of radioactivity reflects dissociation of the dsRBD–R1.1[WC] RNA complex during electrophoresis. Lane 1, no dsRBD; lane 2, no EB; lane 3, 4 µM EB; lane 4, 12 µM EB; lane 5, 20 µM EB; lane 6, 40 µM EB; lane 7, 100 µM EB.