Figure 5

In vitro cleavage of the targeted RNA by M1GS ribozymes. No ribozyme was added to the reaction mixture in lane 1; 10 nM of M1GS-HCV/C₆₇ (lanes 2), M1GS-HCV/C₆₇* (lanes 3) and Chol-M1GS/C₆₇ (lane 4) were incubated with ³²P-labeled substrate RNA (S_1–584, 10 nM) at 37°C in a volume of 10 μl for 30 min in cleavage buffer. As a control, equal volumes of M1GS-HCV/C₆₇ and Chol-M1GS-HCV/C₆₇ were separately incubated with another ³²P-labeled substrate RNA (HCMV UL97 RNA) in the same condition (lanes 5 and 6) Cleavage products were separated on 15% polyacrylamide gels containing 8 M urea. The RNA makers were transcribed in vitro by T7 RNA polymerase with DNA templates linearized by different restriction enzymes (lane M).