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Aim: To study the preparation and cleavage activity of Rz1196 directed against HPV-6bE1 and HPV-11E1 (HPV-6b/11E1) transcripts in vitro. Methods: HPV-6b/11E1 gene fragments were cloned into T-vector under the control ofT7 promoter, ～(32)P-labeted HPV-6b/11E1 transcripts as target-RNAs were transcribed in vitro and purified by PAGE.Rz1198 gene designed as a universal ribozyme for both HPV-6b/11E1 transcripts was cloned into vector p1.5 between5’-cis-Rz and 3’-cis-Rz. a2P-labeled Rzl198 transcript was gel-purified, incubated with target-RNAs at different condi-tions and autoradiographed after denaturing gel-electrophoresis. Results: Rz1198 was active at 37℃. The optimaltemperature was 50℃. For HPV-6bE1, k_m = 12.2 nmol/L, k_cat = 0.18 min～(-1); For HPV-11E1, k_m= 14.7 nmol/L,k_cat =0.14 min～(-1). All these revealed that the design of Rz1198 was correct. It could be a universal ribozyme for thetwo substrates--HPV-6bE1 and HPV-11E1 transcripts. Conclusion: Rz1198 prepared in vitro possesses the perfect
Aim: To study the preparation and cleavage activity of Rz1196 directed against HPV-6bE1 and HPV-11E1 (HPV-6b / 11E1) transcripts in vitro. Methods: HPV-6b / 11E1 gene fragments were cloned into T-vector under the control of T7 promoter, ~ (32) P-labeted HPV-6b / 11E1 transcripts as target-RNAs were transcribed in vitro and purified by PAGE. Rz1198 gene designed as a universal ribozyme for both HPV-6b / 11E1 transcripts was cloned into vector p1.5 between5’-cis-Rz and 3’-cis-Rz. a2P-labeled Rzl198 transcript was gel-purified, incubated with target-RNAs at different condiments and autoradiographed after denaturing gel-electrophoresis. . For HPV-6bE1, k_m = 12.2 nmol / L and k_cat = 0.18 min -1, For HPV-11E1, k_m = 14.7 nmol / L and k_cat = 0.14 min -1, . All these revealed that the design of Rz1198 was correct. It could be a universal ribozyme for the two substrates - HPV-6bE1 and HPV-11E1 transcripts. Conclusion: Rz1198 prepared in vitr o possesses the perfect