Recombination-mediated PCR-directed plasmid construction in vivo in yeast
Recombination-mediated PCR-directed plasmid construction in vivo in yeast
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DOI：
10.1093/nar/25.2.451
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发表时间：
1997-01-15
影响因子：
14.9
通讯作者：
Paddon, C
中科院分区：
文献类型：
--
作者：
Oldenburg, KR;Vo, KT;Paddon, C
我们将聚合酶链式反应定向重组技术扩展到酿酒酵母中，以开发一种在缺乏合适的限制性内切酶的情况下构建质粒或基因的简单方法。待克隆的DNA经PCR扩增，与线性化酵母质粒有30-40个碱基的同源性。共转化到酵母中，导致了由PCR寡核苷酸控制的同源重组ATA位置。
We have extended the technique of PCR-directed recombination in Saccharomyces cerevisiae to develop a simple method for plasmid or gene construction in the absence of suitable restriction sites. The DNA to be cloned is PCR-amplified with 30-40 bp of homology to a linearized yeast plasmid. Co-transformation into yeast results in homologous recombination ata position directed by the PCR oligonucleotides.