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COSPLAY: An expandable toolbox for combinatorial and swift generation of expression plasmids in yeast

机译:COSPLAY:可扩展的工具箱,用于组合和快速生成酵母中的表达质粒

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摘要

A large number of genetic studies in yeast rely on the use of expression vectors. To facilitate the experimental approach of these studies, several collections of expression vectors have been generated (YXplac, pRS series, etc.). Subsequently, these collections have been expanded by adding more diversity to many of the plasmid features, including new selection markers and new promoter sequences. However, the ever growing number of plasmid features makes it unrealistic for research labs to maintain an up-to-date collection of plasmids. Here, we developed the COSPLAY toolbox: a Golden Gate approach based on the scheme of a simple modular plasmid that recapitulates and completes all the properties of the pRS plasmids. The COSPLAY toolbox contains a basal collection of individual functional modules. Moreover, we standardized a simple and rapid, software-assisted protocol which facilitates the addition of new personalized modules. Finally, our toolbox includes the possibility to select a genomic target location and to perform a single copy integration of the expression vector.
机译:酵母中的大量遗传研究都依赖于表达载体的使用。为了促进这些研究的实验方法,已经产生了表达载体的若干集合(YXplac,pRS系列等)。随后,通过向许多质粒特征(包括新的选择标记和新的启动子序列)添加更多的多样性来扩展这些集合。但是,质粒功能的不断增长使研究实验室无法保持质粒的最新收集是不现实的。在这里,我们开发了COSPLAY工具箱:基于简单模块化质粒方案的Golden Gate方法,该方案概括并完善了pRS质粒的所有特性。 COSPLAY工具箱包含各个功能模块的基础集合。此外,我们标准化了一个简单,快速,软件辅助的协议,该协议有助于添加新的个性化模块。最后,我们的工具箱包括选择基因组目标位置并执行表达载体的单拷贝整合的可能性。

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