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PCR介导的重组与诱变。通过重叠延伸拼接定制基因。

PCR-mediated recombination and mutagenesis. SOEing together tailor-made genes.

作者信息

Horton R M

机构信息

Department of Biochemistry, College of Biological Sciences, St. Paul, MN 55108, USA.

出版信息

Mol Biotechnol. 1995 Apr;3(2):93-9. doi: 10.1007/BF02789105.

Abstract

Gene Splicing by Overlap Extension (gene SOEing) is a sequence-independent method for site-directed mutagenesis and/or recombination of DNA molecules. It is based on the idea that a PCR product can be engineered by adding or changing sequences at its ends so that the product can itself be used to prime DNA synthesis in a subsequent overlap-extension reaction to create mutant or recombinant molecules. As the engineered genes are created in vitro without reliance on host organisms or restriction sites, gene SOEing provides a powerful and versatile tool for genetic investigation and engineering.

摘要

重叠延伸基因剪接(基因SOEing)是一种用于DNA分子定点诱变和/或重组的与序列无关的方法。它基于这样一种理念,即通过在PCR产物的末端添加或改变序列来对其进行改造,以便该产物本身可用于在随后的重叠延伸反应中引发DNA合成,从而创建突变或重组分子。由于改造后的基因是在体外创建的,无需依赖宿主生物体或限制位点,基因SOEing为基因研究和工程提供了一种强大且通用的工具。

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