Mutation detection with MutH, MutL, and MutS mismatch repair proteins
Mutation detection with MutH, MutL, and MutS mismatch repair proteins
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DOI：
10.1073/pnas.93.9.4374
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发表时间：
1996-04-30
影响因子：
11.1
通讯作者：
Modrich, P
中科院分区：
文献类型：
--
作者：
Smith, J;Modrich, P
大肠杆菌甲基介导的错配修复是由MutH核酸内切酶的MutS-、MutL-和ATP-依赖性激活启动的，MutH核酸内切酶在错配附近的d（GATC）位点裂解。该反应提供了一种有效的方法，用于检测PCR扩增后通过遗传上不同的序列杂交产生的异源双链体中的错配。已在PCR异源双链体中检测到转换和颠换突变以及一个、两个和三个核苷酸插入/缺失突变体的多个实例，其大小范围为400 bp至2.5 kb。同源双链体的背景切割主要是由于扩增过程中发生的聚合酶错误，MutHLS反应提供了PCR过程中出现的突变序列发生率的估计。
Escherichia coli methyl-directed mismatch repair is initiated by MutS-, MutL-, and ATP-dependent activation of MutH endonuclease, which cleaves at d(GATC) sites in the vicinity of a mismatch. This reaction provides an efficient method for detection of mismatches in heteroduplexes produced by hybridization of genetically distinct sequences after PCR amplification. Multiple examples of transition and transversion mutations, as well as one, two, and three nucleotide insertion/deletion mutants, have been detected in PCR heteroduplexes ranging in size from 400 bp to 2.5 kb. Background cleavage of homoduplexes is largely due to polymerase errors that occur during amplification, and the MutHLS reaction provides an estimate of the incidence of mutant sequences that arise during PCR.