Efficient targeted integration at leu1-32 and ura4-294 in Schizosaccharomyces pombe.
Efficient targeted integration at leu1-32 and ura4-294 in Schizosaccharomyces pombe.
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作者：
J. Keeney;J. Boeke
同源整合到裂殖酵母中的特性尚未得到很好的描述。在这项研究中，我们检测了携带leu1+和ura4+基因的质粒整合到它们的染色体上。基因组DNA印迹分析表明，大多数转化子通过同源重组整合了一个或多个拷贝的质粒载体，但转化为Leu1+或ura4+的基因比例要小得多。两个基因均未观察到非同源重组事件。我们描述了用于在S.pombe的leu1-32和ura4-294基因座进行靶向整合的通用的leu1+和ura4+质粒的构建。
Homologous integration into the fission yeast Schizosaccharomyces pombe has not been well characterized. In this study, we have examined integration of plasmids carrying the leu1+ and ura4+ genes into their chromosomal loci. Genomic DNA blot analysis demonstrated that the majority of transformants have one or more copies of the plasmid vector integrated via homologous recombination with a much smaller fraction of gene conversion to leu1+ or ura4+. Non-homologous recombination events were not observed for either gene. We describe the construction of generally useful leu1+ and ura4+ plasmids for targeted integration at the leu1-32 and ura4-294 loci of S. pombe.