Laboratory of Biology and Information Science, School of Life Sciences, East China Normal University, Shanghai 200062, China.
Int J Mol Sci. 2023 Feb 7;24(4):3243. doi: 10.3390/ijms24043243.
Cell morphology is an essential and phenotypic trait that can be easily tracked during adaptation and evolution to environmental changes. Thanks to the rapid development of quantitative analytical techniques for large populations of cells based on their optical properties, morphology can be easily determined and tracked during experimental evolution. Furthermore, the directed evolution of new culturable morphological phenotypes can find use in synthetic biology to refine fermentation processes. It remains unknown whether and how fast we can obtain a stable mutant with distinct morphologies using fluorescence-activated cell sorting (FACS)-directed experimental evolution. Taking advantage of FACS and imaging flow cytometry (IFC), we direct the experimental evolution of the population undergoing continuous passage of sorted cells with specific optical properties. After ten rounds of sorting and culturing, a lineage with large cells resulting from incomplete closure of the division ring was obtained. Genome sequencing highlighted a stop-gain mutation in , leading to a dysfunctional AmiC division protein. The combination of FACS-based selection with IFC analysis to track the evolution of the bacteria population in real-time holds promise to rapidly select and culture new morphologies and association tendencies with many potential applications.
细胞形态是一种重要的表型特征，在适应和进化过程中很容易追踪到环境变化。由于基于细胞光学特性的大量细胞的定量分析技术的快速发展，在实验进化过程中可以很容易地确定和跟踪形态。此外，新的可培养形态表型的定向进化可以在合成生物学中找到用途，以改进发酵过程。目前尚不清楚我们是否以及如何能够快速获得具有独特形态的稳定突变体，使用荧光激活细胞分选 (FACS) 定向实验进化。我们利用 FACS 和成像流式细胞术 (IFC)，对经历具有特定光学特性的分选细胞连续传代的群体进行实验进化。经过十轮分选和培养，获得了由于分裂环不完全闭合而导致的大细胞系。基因组测序突出了一个导致 AmiC 分裂蛋白功能失调的无义突变。基于 FACS 的选择与 IFC 分析相结合，实时跟踪细菌群体的进化，有望快速选择和培养新的形态以及与许多潜在应用相关的关联趋势。
