Pacific Northwest Research Institute, Seattle, Washington, USA.
Yeast. 2022 Jun;39(6-7):354-362. doi: 10.1002/yea.3802. Epub 2022 Jun 23.
Meiotic mapping, a linkage-based method for analyzing the recombinant progeny of a cross, has long been a cornerstone of genetic research. The yeast Saccharomyces cerevisiae is a powerful system because it is possible to isolate and cultivate the four products (spores) of a single meiotic event. However, the throughput of this process has historically been limited by the process of identifying tetrads in a heterogeneous population of vegetative cells, tetrads, and dyads followed by manual separation (dissection) of the spores contained in a tetrad. To date, methods that facilitate high throughput characterization and isolation of meiotic progeny have relied on genetic engineering. Here, we characterize the ability of the fluorescent dye DiBAC (5) to stain yeast tetrads and dyads as well as to adhere to spores following bulk tetrad disruption. Applications include quantitative assays of sporulation rates and efficiency by flow cytometry as well as enrichment of intact tetrads, dyads, or disrupted spores by fluorescence-activated cell sorting in strains that have not been genetically modified.
减数分裂作图是一种基于连锁分析的方法，用于分析杂交后代的重组子，长期以来一直是遗传研究的基石。酵母酿酒酵母是一种强大的系统，因为可以分离和培养单个减数分裂事件的四个产物（孢子）。然而，由于需要鉴定异质群体中的四分体、四分体和二联体，然后手动分离（解剖）四分体中的孢子，因此该过程的通量在历史上一直受到限制。迄今为止，促进高通量鉴定和分离减数分裂后代的方法依赖于遗传工程。在这里，我们描述了荧光染料 DiBAC（5）对酵母四分体和二联体的染色能力，以及在大量四分体破坏后对孢子的附着能力。应用包括通过流式细胞术定量测定孢子形成率和效率，以及通过荧光激活细胞分选对未遗传修饰的菌株中的完整四分体、二联体或破坏的孢子进行富集。
