Several microbes are polyploid, meaning they contain several copies of their chromosome. Cyanobacteria, while holding great potential as photosynthetic cell factories of various products, are found among them. In these clades the diversity of genetic elements that serve within the basic molecular toolbox is often limiting. To assist mining for the latter, we present here a method for the generation of fully segregated genomic libraries, specifically designed for polyploids. We provide proof-of-principle for this method by generating a fully segregated genomic promoter library in the cyanobacterium Synechocystis sp. PCC 6803. This new tool was first analyzed through fluorescence activated cell sorting (FACS) and then a fraction was further characterized regarding promoter sequence. The location of libraries on the chromosome provides a better reflection of the behavior of its elements. Our work presents the first method for constructing fully segregated genomic libraries in polyploids, which may facilitate their usage in synthetic biology applications.

中文翻译：

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多倍体生物（如集胞藻）完全分离的基因组文库的构建。PCC 6803

一些微生物是多倍体，这意味着它们含有多个染色体拷贝。其中蓝藻细菌作为各种产品的光合细胞工厂具有巨大的潜力。在这些进化枝中，基本分子工具箱中发挥作用的遗传元件的多样性通常是有限的。为了帮助挖掘后者，我们在这里提出了一种专门为多倍体设计的完全分离的基因组文库的生成方法。我们通过在蓝藻集胞藻属中生成完全分离的基因组启动子文库，为该方法提供了原理验证。PCC 6803。首先通过荧光激活细胞分选 (FACS) 对这一新工具进行分析，然后进一步对部分启动子序列进行表征。文库在染色体上的位置可以更好地反映其元素的行为。我们的工作提出了第一种在多倍体中构建完全分离的基因组文库的方法，这可能有助于它们在合成生物学应用中的使用。