【摘要】 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。更多还原

【Abstract】 The genes of hammerhead ribozymes targeting against two and three sites on negative strand of potato spindle tuber viroid (PSTVd) were designed, synthesized and cloned according to the action manner of hammerhead ribozyme. After transcription in vitro , the PSTVd negative RNA was mixed with the ribozyme containing two or three cleavage sites separately and incubated two hours at 37℃. The results showed that the ribozymes containing two and three cleavage sites have highly catalytic activity, and the size of the cleft products and released by the ribozyme containing two cleavage sites is correspond with the expected value. Although the ribozyme containing three cleavage sites has highly catalytic activity, the ribozyme is effective only for one cleavage site. The applied prospect for two ribozymes was discussed in the paper.更多还原