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根据锤头型核酶的作用模式 ,设计、合成并克隆了特异切割马铃薯纺锤形块茎类病毒 (PSTVd)负链RNA不同区域位点的双价和三价锤头型核酶基因。通过体外转录 ,将PSTVd负链RNA分别与双价和三价核酶混合 ,37℃温育 2h。结果表明 ,双价核酶和三价核酶均表现出较高的切割活性 ,其中双价核酶处理的切割产物的大小与理论值相符合。三价核酶虽表现出较高的切割活性 ,但只是其中一价核酶在起作用。讨论了二价和三价核酶的应用前景。
According to the mode of action of hammerhead ribozyme, bivalent and trivalent hammerhead ribozyme genes were designed, synthesized and cloned specifically to different regions of negative strand RNA of potato spindle-shaped tuberovirus (PSTVd). By in vitro transcription, the PSTVd minus-strand RNAs were mixed with the bivalent and trivalent ribozymes, respectively, and incubated at 37 ° C for 2h. The results showed that both divalent and trivalent ribozymes exhibited higher cleavage activity, in which the size of the divalent ribozyme-treated cleavage product was consistent with the theoretical value. Trivalent ribozyme, although exhibiting high cleavage activity, is only one of the monovalent ribozymes at work. Discuss the application prospect of bivalent and trivalent ribozyme.