Figure 2. Cleavage in vitro of the mglB mRNA by the RNase P holoenzyme in the absence or presence of stem EGSs.
A. Activities of stem EGS155 and EGS155R. B. Activities of stem EGS52, EGS73, and EGS148. If needed, 10 nM M1 RNA and 100 nM C5 protein in buffer PA were added to reconstitute the RNase P holoenzyme. The mglB mRNA cleavage products were separated on 8% polyacrylamide/7 M urea gels along with the mglB mRNA alone or the mglB mRNA with the RNase P holoenzyme. Internally labeled pSupS1 ptRNA was used as a positive control to check for activity of the reconstituted RNase P holoenzyme. EGSs were added in 100-, 50-, and 10-fold molar excess to the mglB mRNA, denoted by black triangles.
