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1. 发明公开

KR1020090036452A 인버스 ＰＣＲ을 이용한 부위-지정 돌연변이 유발방법 无效
标题翻译：使用反向PCR的位点指导性突变的方法
公开(公告)号：KR1020090036452A
公开(公告)日：2009-04-14
申请号：KR1020070101650
申请日：2007-10-09
申请人： 주식회사 엔지노믹스
发明人： 추승호 , 이병철 , 김선혜 , 이경민 , 류재준 , 이민영 , 이미혜 , 류기혁
IPC分类号： C12N15/09 , C12N15/00 , C12N15/10
摘要： A method for inducing site-directed mutagenesis by using inverse PCR is provided to change, remove or add a specific base of a gene, promoter or other DNA pieces which are cloned in a vector by using inverse PCR and a mixture of 3 kinds of enzymes. A method for inducing site-directed mutagenesis by using inverse PCR comprises the steps of: (i) performing inverse PCR by using a primer pair of the opposite direction having mutagenesis at the desired site with the circular double strand DNA molecule as a mold; and (ii) reaction of 3 kinds of enzyme by adding a mixture of 3 kinds of enzymes to PCR products. The 3 kinds of enzyme comprises kinase for phosphorylating 5 ' end of a primer; ligase for connecting both ends of the amplified PCR products and a restriction enzyme for cutting the methylated template strand. The mutagenesis is point mutagenesis(substitution), deletion or insertion.
摘要翻译：提供使用反向PCR诱导定点诱变的方法，通过使用反向PCR和3种酶的混合物来改变，去除或添加克隆在载体中的基因，启动子或其他DNA片段的特异性碱基。通过使用反向PCR诱导定点诱变的方法包括以下步骤：（i）通过使用具有在所需位点的诱变的相反方向的引物对以圆形双链DNA分子作为模具进行逆PCR; 和（ii）通过向PCR产物中加入3种酶的混合物来反应3种酶。 3种酶包括用于磷酸化底物5'末端的激酶; 连接酶用于连接扩增的PCR产物的两端和用于切割甲基化模板链的限制酶。诱变是点诱变（取代），缺失或插入。

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