In vitro cleavage of HPV16 E6 and E7 RNA fragments by synthetic ribozymes and transcribed ribozymes from RNA‐trimming plasmids

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Abstract
A RNA‐trimming plasmid pRG523 is constructed, in which three Rz genes, GR5(5'‐cis‐Rz gene), HR2G(trans‐Rz gene) and GR3(3'‐cis‐Rz gene), are arranged in the order from 5' to 3' downstream from the T7 promoter. In vitro transcription of this plasmid shows that the trans‐Rz can be trimmed to definite lengths by the cis‐Rz on both sides of the trans‐Rz. In vitro cleavage of HPV16 E6 and E7 RNA fragments of different lengths by synthetic Rz and that of E7 RNA with a length of 171 nt by synthetic Rz and transcribed Rzs with different lengths of flanking sequences is studied. The results show that the non‐base‐pairing flanking sequences on both Rz and target RNA can affect the cleavage reaction.