A RAPID METHOD FOR LOCALIZED MUTAGENESIS OF YEAST GENES
A RAPID METHOD FOR LOCALIZED MUTAGENESIS OF YEAST GENES
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DOI：
10.1002/yea.320080202
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发表时间：
1992-02-01
期刊：
影响因子：
2.6
通讯作者：
PARKER, R
中科院分区：
文献类型：
--
作者：
MUHLRAD, D;HUNTER, R;PARKER, R
我们已经开发了一个简单的程序酵母基因的局部诱变。在该技术中，首先在诱变聚合酶链式反应（PCR）条件下扩增感兴趣的区域。PCR产物与含有与PCR产物两端同源性的缺口质粒的共转化允许体内重组，以用诱变的DNA修复差距。该方法是有效的，允许靶向特定区域进行诱变，并且不需要在大肠杆菌中进行亚克隆步骤。
We have developed a simple procedure for the localized mutagenesis of yeast genes. In this technique the region of interest is first amplified under mutagenic polymerase chain reaction (PCR) conditions. Cotransformation of the PCR product with a gapped plasmid containing homology to both ends of the PCR product allows in vivo recombination to repair the gap with the mutagenized DNA. This procedure is efficient, allows targeting of specific regions for mutagenesis, and requires no subcloning steps in Escherichia coli.