SITE-SPECIFIC DELETION OF THE N-TERMINAL SEGMENTS FROM EcoRI ENDONUCLEASE USING THE POLYMERASE CHAIN REACTION

作     者：杨香娇 陈常庆 王德宝 杨胜利 

作者机构：Shanghai Institute of Biochemistry Academia Sinica Shanghai 200031 PRC Shanghai Center of Biotechnology Academia Sinica Shanghai 200031 PRC 

出 版 物：《Science China Chemistry》 (中国科学（化学英文版）)

年 卷 期：1991年第34卷第12期

页      面：1436-1443页

核心收录：

学科分类：0810[工学-信息与通信工程] 0830[工学-环境科学与工程（可授工学、理学、农学学位）] 07[理学] 0708[理学-地球物理学] 0805[工学-材料科学与工程（可授工学、理学学位）] 0703[理学-化学] 0704[理学-天文学] 0812[工学-计算机科学与技术（可授工学、理学学位）] 

主　　题：PCR site-specific deletion EcoRI endonuclease. 

摘      要：We developed a general and efficient method for directing the deletions of DNA sequences of any lengths using polymerase chain reaction (PCR). The method was based on in vitro amplification of target sequences with site-specific deletions, Klenow end-flushing and blunt-end cloning. As an example, it was used to delete the restriction gene encoding EcoRI endonuclease, resulting in plasmids expressing two truncated forms. Assays using SDS-PAGE and gel retardation revealed the important role of the amphipathic helix (29-43) of the EcoRI endonuclease in binding to its cognate substrate.