利用扩展错配引物的寡核苷酸定向诱变诱导多个替代突变

Alex Lichtler , Gordon L. Hager
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引用次数: 2

摘要

我们已经开发了一种称为寡核苷酸扫描诱变的方法,它使用寡核苷酸在一个步骤中突变多达12个连续的碱基。该方法的一些优点是替换突变的位置和序列完全由研究者指定,并且可以很容易地生成突变的组合。该技术使用间隙底物和大肠杆菌坝甲基化纠错机制来增加突变体的产量。
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Induction of multiple replacement mutations by oligonucleotide-directed mutagenesis with extended mismatch primers

We have developed a method called oligo-scanning mutagenesis that uses oligonucleotides to mutate up to 12 contiguous bases in a single step. Some advantages of this procedure are that the position and sequence of the replacement mutations are completely specified by the investigator, and combinations of mutations can easily be generated. The technique uses a gapped substrate and the Escherichia coli dam methylation error-correcting mechanism to increase the yield of mutants.

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Gene targeting in murine embryonic stem cells: Introduction of specific alterations into the mammalian genome A solution hybridization method for quantification of mRNAs: Determining the amount and stability of oncogene mRNA The use of transgenic mice for short-term, in vivo mutagenicity testing Author index volume 7 Subject index volume 7
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