A RAPID METHOD FOR LOCALIZED MUTAGENESIS OF YEAST GENES

被引：413

作者：

MUHLRAD, D ^{[1
]}

HUNTER, R ^{[1
]}

PARKER, R ^{[1
]}

机构：

[1] UNIV ARIZONA, DEPT MOLEC & CELLULAR BIOL, TUCSON, AZ 85721 USA

来源：

YEAST | 1992年 / 8卷 / 02期

关键词：

SITE-DIRECTED MUTAGENESIS; SACCHAROMYCES-CEREVISIAE; PCR;

D O I：

10.1002/yea.320080202

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：79 / 82

页数：4

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