Oligo诱导突变的改进方法

Oligo诱导突变的改进方法 AN IMPROVED METHOD FOR OLIGO DIRECTED MUTAGEMESIS 叶林柏 Ye Linbai 1 first-author 叶璨樱 Ye Chanying 1 郜金荣 Gao Jinrong 1 徐进平 Xu Jinping 1 孟小林 Mong Xiaolin 1 胡敏 Hu Min 1 武汉大学生命科学学院 武汉大学生命科学学院 College of Life Sciences,Wuhan University College of Life Sciences,Wuhan University 介绍一种改进的Ｏｌｉｇｏ诱导ＤＮＡ定点突变方法，此法制备的含Ｕ野生型模板ＤＮＡ在ＪＭ１０７中的存活率从５％～１０％下降至１％～２％；诱变反应终产物转染ＪＭ１０７产生的斑数比原法提高１５～２０倍，诱导连续４个核苷酸替换产生的突变频率达８０％～８５％，即使诱导连续１２个核苷酸的缺失，也能获得４０％～５０％的突变频率． An improved method for oligo directed site specific mutagenesis was developed. The Survivor of u containing template M13 DNA prepared by modified protocol was droped down from 5%￣10% originally to 1%～2% in JM107. The plaques number generated by transfection of JM107 with mutagenic reaction products increase 15 to 20 fold. The frequence of mutation obtained by the improved method was 80%￣85% for 2～4 nucleotidcs alternation and 40%￣50% for 9～12 nucleotides deletion. With this improved method, we can obtain the site specific mutants of genes easily. DNA定点实变 方法 Oligo诱导 DNA site mutation improved methad oligo directed Q78 国家自然科学基金(61672102) 1998-02-01 2021-04-01 2