酵母体内重组介导的PCR定向质粒构建
Recombination-mediated PCR-directed plasmid construction in vivo in yeast.
作者信息
Oldenburg K R, Vo K T, Michaelis S, Paddon C
机构信息
Affymax Research Institute, 4001 Miranda Avenue, Palo Alto, CA 94304, USA.
出版信息
Nucleic Acids Res. 1997 Jan 15;25(2):451-2. doi: 10.1093/nar/25.2.451.
Abstract
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 at a position directed by the PCR oligonucleotides.
摘要
我们已经扩展了酿酒酵母中PCR定向重组技术，以开发一种在缺乏合适限制酶切位点的情况下进行质粒或基因构建的简单方法。待克隆的DNA用与线性化酵母质粒具有30 - 40 bp同源性的引物进行PCR扩增。共转化到酵母中会导致在PCR寡核苷酸引导的位置发生同源重组。
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