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흑조위축병 바이러스 RNA 를 절단하는 망치머리형 라이보자임의 제작
박종석(Jong Sug Park),김주곤(Ju Kon Kim),손성한(Seong Han Sohn),이석순(Sug Soon Lee),황영수(Young Soo Hwang) 한국응용생명화학회 1995 Applied Biological Chemistry (Appl Biol Chem) Vol.38 No.6
To develop an antiviral agent for the rice black-streaked dwarf virus (RBSDV), a hammerhead type ribozyme, which has a potential target site on the genome segment 3, was designed. Oligonucleotides for the ribozyme and its substrate were synthesized, annealed, and cloned into a plasmid pBluescript II KS(+). Ribozyme and substrate RNAs were then synthesized by in vitro transcription with T₃ RNA polymerase, obtaining RNAs in expected size, 193 and 182 nucleotides, respectively. The substrate RNA was efficiently cleaved into two fragments when incubated with the ribozyme at 55℃, while the cleavage was not detected at 37℃. In addition, the segment 3 RNA of RBSDV was also cleaved into two fragments by the same ribozyme at 55℃. Taken together, our results demonstrated that the hammerhead ribozyme has an in vitro endonucleolytic activity and may be used as an antiviral agent in transgenic plants.