Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Ontario, Canada.
Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada.
Yeast. 2024 Oct;41(10):585-592. doi: 10.1002/yea.3978. Epub 2024 Sep 9.
Common Saccharomyces cerevisiae lab yeast strains derived from S288C have meiotic defects and therefore are poor sporulators. Here, we developed a plasmid system containing corrected alleles of the MKT1 and RME1 genes to rescue the meiotic defects and show that standard BY4741 and BY4742 strains containing the plasmid display faster and more efficient sporulation. The plasmid, pSPObooster, can be maintained as an episome and easily cured or stably integrated into the genome at a single locus. We demonstrate the use of pSPObooster in low- and high-throughput yeast genetic manipulations and show that it can expedite both procedures without impacting strain behavior.
常见的酿酒酵母实验室酵母菌株源自 S288C，具有减数分裂缺陷，因此是不良的孢子形成体。在这里，我们开发了一种含有 MKT1 和 RME1 基因校正等位基因的质粒系统，以挽救减数分裂缺陷，并表明含有质粒的标准 BY4741 和 BY4742 菌株显示出更快、更有效的孢子形成。质粒 pSPObooster 可以作为附加体维持，并且可以很容易地在单个基因座处被消除或稳定整合到基因组中。我们展示了 pSPObooster 在低通量和高通量酵母遗传操作中的用途，并表明它可以在不影响菌株行为的情况下加速这两个过程。
