摘要
通过微机对ｂｃｌ－２ＲＮＡ二级结构的分析，设计针对ｂｃｌ－２片段５＇ＣＧＣＧＡＣＣＣＧＧＵＣＧＣＣＡＧＧＡＣＣＵＣＧ３＇的“锤头状”（Ｈａｍｍｅｒｈｅａｄ）核酶（Ｒｉｂｏｚｙｍｅ，ＲＤ）基因，平端连接于ｐＧＥＭ－３Ｚｆ（－）ＨｉｎｃⅡ位点，克隆后经测序表明序列正确，ｂｃｌ－２和Ｒｉｂｏｚｙｍｅ基因经体外转录，５０℃作用２ｈ，从１６５６－１６５７（Ｃ－Ｇ）位之间切断ｂｃｌ－２ＲＮＡ片段．
After analysis of the secondary structure of bcl-2 RNA by computer,a hammerhead ribozyme have been designed to cleave the sequence 5'CGC GAC CCG GUC GCC AGG ACC UCG 3' of bcl-2 mRNA. The DNA of ribozyme was cloned into the Hinc Ⅱ site of pGEM-3Zf (-), named as 3ZRZ (+)/ (-), sequenced correctly and transcribed in vitro. The transcripts were mixed at the condition of 15 mmol/L MgCl2 , 50℃, for 2 h. The bcl-2 transcripts were cleaved by ribozyme at the site between 1656 and 1657 of bcl-2 RNA, demonstrating strong cleavage activity.
基金
国家自然科学基金
