" Study on the cleavage of potato lea/roll virus cost protein gene in vitro by ribozyme
Abstract: A hammerhead structure ribozyme was designed and synthesized to potato leafroll virus Chinese isolate (PLRV - Ch) genome within 356 to 358 nt "GUC" in coat protein (CP) gene. DNA sequence encoding the ribozyme was inserted into the position downstream from SP6 promoter of in vitro transcription vector pSPT19. The target PLRV CP gene cDNA was subcloned in vector pSPT18. The RNA molecules of riboZyme and target Sequence were generated by transcription with SP6 RNA polymerase in vitro. After cleavage reaction at 41℃, two.expected short RNA fragments were observed.