【摘要】 在锤头型核酶下游增加一段核酶作用的靶序列 ,使之成为自切割的核酶 .将自切割核酶基因合成 ,扩增并克隆在质粒pBluescriptSK上 ,经连续 4次克隆获得含 10个拷贝的自切割核酶基因的载体 .5%聚丙烯酰胺变性凝胶电泳结果显示 :体外转录过程中多体酶发生了自切割 ,核酶转录物切割成从 70nt到 70 6nt的RNA等梯度带 ,因此 ,可以作为RNA分子质量标记之用更多还原

【Abstract】 A self cleaving ribozyme was comprised of a hammerhead ribozyme and its target sequence located the downstream of the hammerhead ribozyme . The self cleaving ribozyme gene was synthesized , amplified and cloned into the Bluescript SK plasmid. The construct harboring 10 copies of the self cleaving ribozyme gene was obtained through successively cloning for four times. The result of polyacrylamid denatured gel electrophoresis showed: the multimeric ribozymes caused self cleaving during the transcription reaction in vitro and formed RNA step ladders from 70 nt to 706 nt, which indicates that the self cleavage ribozyme transcripts can be used as RNA molecular mass markers.更多还原