Modules for cloning-free chromatin tagging in Saccharomyces cerevisae
Modules for cloning-free chromatin tagging in Saccharomyces cerevisae
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DOI：
10.1002/yea.1580
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发表时间：
2008-03-01
期刊：
影响因子：
2.6
通讯作者：
Meister, Peter
中科院分区：
文献类型：
--
作者：
Rohner, Sabine;Gasser, Susan M.;Meister, Peter
我们描述了一个简单的两步PCR为基础的方法插入阵列的乳糖或泰特运营商（lacO或tetO）在特定的基因座在芽殖酵母基因组。该方法需要用识别感兴趣的基因座的经典长引物插入通过PCR产生的标记，然后用带有lacI或tetR结合基序阵列的线性化质粒替换该标记。使用这种技术，位于酵母基因组或酵母人工染色体上的基因座可以被有效地标记。我们提供了一组具有不同标记的质粒，用于将lacO或tetO重复序列无克隆整合到酵母基因组中。版权所有（c）2008约翰威利父子有限公司。
We describe a straightforward two-step PCR-based method to insert arrays of lac or tet operators (lacO or tetO) at specific loci in the budding yeast genome. The method entails insertion of a marker generated by PCR with classical long primers recognizing the locus of interest, followed by the replacement of this marker by a linearized plasmid bearing an array of lacI- or tetR-binding motifs. Using this technique, loci located either in the yeast genome or on yeast artificial chromosomes can be efficiently tagged. We provide a set of plasmids with different markers for cloning-free integration of lacO or tetO repeats into the yeast genome. Copyright (c) 2008 John Wiley & Sons, Ltd.