Centromeres (CENs) are the chromosomal regions promoting kinetochore formation for faithful chromosome segregation. In yeasts, CENs have been recognized as the essential elements for extra-chromosomal DNA stabilization. However, the epigeneticity of CENs makes their localization on individual chromosomes very challenging, especially in many not well-studied nonconventional yeast species. Previously, we applied a stepwise method to identify a 500-bp CEN5 from Scheffersomyces stipitis chromosome 5 and experimentally confirmed its critical role on improving plasmid stability. Here we report a library-based strategy that integrates in silico GC₃ chromosome scanning and high-throughput functional screening, which enabled the isolation of all eight S. stipitis centromeres with a 16 000-fold reduction in sequence very efficiently. Further identification of a 125-bp CEN core sequence that appears multiple times on each chromosome but all in the unique signature GC₃-valley indicates that CEN location might be accurately discerned by their local GC₃ percentages in a subgroup of yeasts.

中文翻译：

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快速分离裂殖酵母中的着丝粒

着丝粒（CENs）是促进线粒体形成的染色体区域，用于忠实的染色体分离。在酵母中，CENs被认为是染色体外DNA稳定的必要元素。然而，CENs的表观遗传学使得它们在单个染色体上的定位非常具有挑战性，尤其是在许多未经充分研究的非常规酵母物种中。以前，我们应用了一种逐步方法，从裂殖酵母5号染色体上鉴定了一个500 bp的CEN5，并通过实验证实了其对提高质粒稳定性的关键作用。在这里，我们报告基于库的战略集成在硅片GC ₃号染色体的扫描和高通量筛选的功能，这使所有八个的隔离小号。的拟南芥着丝粒，序列减少了16000倍，非常有效。进一步鉴定出在每个染色体上多次出现但都在唯一签名GC _3-谷中出现的125 bp CEN核心序列，表明可以通过它们在酵母亚组中的局部GC ₃百分比准确地识别CEN位置。