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通过聚乙二醇/醋酸锂转化和测序对酿酒酵母进行适应性实验室进化的方案。

Protocol for adaptive laboratory evolution of S. cerevisiae by PEG/LiAc transformation and sequencing.

作者信息

Jiang Bei, Xiao Chuyao, Liu Li

机构信息

MOE Key Laboratory of Gene Function and Regulation, State Key Laboratory of Biocontrol, Innovation Center for Evolutionary Synthetic Biology, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China; Department of Health Technology and Informatics, The Hong Kong Polytechnic University, Hong Kong SAR, China.

MOE Key Laboratory of Gene Function and Regulation, State Key Laboratory of Biocontrol, Innovation Center for Evolutionary Synthetic Biology, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China; Greater Bay Area Institute of Precision Medicine (Guangzhou), Fudan University, Nansha District, Guangzhou 511400, China; Institute of Life Sciences, Fudan University, Shanghai 200433, China.

出版信息

STAR Protoc. 2025 Jun 20;6(2):103857. doi: 10.1016/j.xpro.2025.103857. Epub 2025 May 31.

Abstract

Gene deletions perturb cellular homeostasis, affecting gene expression and phenotypes. Here, we present a protocol for serial transfer experiments in Saccharomyces cerevisiae strains following targeted gene knockouts. We describe steps for gene deletion, serial passage optimization, and growth kinetics analysis. We then detail procedures for genomic and transcriptomic profiling. This protocol enables the investigation of changes in yeast strains resulting from gene loss. For complete details on the use and execution of this protocol, please refer to Jiang et al..

摘要

基因缺失会扰乱细胞内稳态,影响基因表达和表型。在此,我们介绍一种在酿酒酵母菌株中进行靶向基因敲除后进行连续传代实验的方案。我们描述了基因缺失、连续传代优化和生长动力学分析的步骤。然后详细说明了基因组和转录组分析的程序。该方案能够研究基因缺失导致的酵母菌株变化。有关该方案的使用和执行的完整详细信息,请参考Jiang等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6990/12163137/b218553ab58a/fx1.jpg

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