Anti-HBV hairpin ribozyme-mediated cleavage of target RNA in vitro

Figure 1 Structure of HpRz plus 5’-cis-Rz and 3’-cis-Rz.

Figure 2 The analysis of HpRz digested by EcoRI and HindIII(20 g·L^-1 agrose gel). lane 1:λ DNA/HindIII marker; lane 2: DL2000 DNA marker; lane 3-7: HpRz digested by EcoRI and HindIII.

Figure 3 in vitro transcript of pHpRZ. HpRz is 83nt. 5’-cis-Rz is 63nt, 3’-cis-Rz is 50nt.

Figure 4 Time course. Specific cleavage of HBV RNA target molecules by HpRz in vitro. Lane 1:substrate control; lane 2:incubated for 6 min; lane 3: 10 min; lane 4:20 min; lane 5:40 min; lane 6:60 min; lane 7:90 min.

Figure 5 Time curves of cleavage reactions of HpRz prepared in vitro.

Figure 6 Lineweaver-burk kinetic plots of the specific cleavage of HBV RNA target molecules by HpRz prepared in vitro. HpRz concentration is 5 nmol·^-1, substate concentration is 80,40,20,10 5 nmol·L^-1. Reaction is at 37 °C for 40 min.