一组大肠杆菌中的lacZ突变，可实现对六种碱基替换中每一种的快速检测。
A set of lacZ mutations in Escherichia coli that allow rapid detection of each of the six base substitutions.
作者信息
Cupples C G, Miller J H
机构信息
Molecular Biology Institute, University of California, Los Angeles 90024.
出版信息
Proc Natl Acad Sci U S A. 1989 Jul;86(14):5345-9. doi: 10.1073/pnas.86.14.5345.
Abstract
We describe the construction of six strains of Escherichia coli with different mutations at the same coding position in the lacZ gene, which specifies the active site glutamic acid residue at position 461 in beta'-galactosidase. Each strain is Lac- and reverts to Lac+ only by restoring the glutamic acid codon. The strains have been designed so that each reverts via one of the six base substitutions. The set of strains allows detection of each transition and transversion simply by monitoring the Lac- to Lac+ frequency, as demonstrated here with characterized mutagens and mutator alleles. These strains are useful for rapidly determining the mutagenic specificity of mutagens at a single site, for detecting low levels of stimulation of certain base substitutions, for monitoring specific base changes in response to various experimental conditions or strain backgrounds, and for isolating new mutator strains.
摘要
我们描述了在lacZ基因相同编码位置具有不同突变的六种大肠杆菌菌株的构建，该基因指定了β-半乳糖苷酶中461位的活性位点谷氨酸残基。每个菌株都是Lac-，只有通过恢复谷氨酸密码子才能回复为Lac+。设计这些菌株的目的是使每个菌株通过六个碱基替换之一进行回复。通过监测Lac-到Lac+的频率，这组菌株能够简单地检测每种转换和颠换，本文用特征性诱变剂和诱变等位基因进行了证明。这些菌株可用于快速确定诱变剂在单个位点的诱变特异性，检测某些碱基替换的低水平刺激，监测响应各种实验条件或菌株背景的特定碱基变化，以及分离新的诱变菌株。
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