Harnessing Single Cell Sorting to Identify Cell Division Genes and Regulators in Bacteria

Cell division is an essential cellular process that requires an array of known and unknown proteins for its spatial and temporal regulation. Here we develop a novel, high-throughput screening method for the identification of bacterial cell division genes and regulators. The method combines the over-expression of a shotgun genomic expression library to perturb the cell division process with high-throughput flow cytometry sorting to screen many thousands of clones. Using this approach, we recovered clones with a filamentous morphology for the model bacterium, Escherichia coli. Genetic analysis revealed that our screen identified both known cell division genes, and genes that have not previously been identified to be involved in cell division. This novel screening strategy is applicable to a wide range of organisms, including pathogenic bacteria, where cell division genes and regulators are attractive drug targets for antibiotic development.

细胞分裂是核心细胞生命过程，其时空调控依赖于一系列已知与未知蛋白质的协同作用。本研究开发了一种全新的高通量筛选方法，用于鉴定细菌细胞分裂相关基因及其调控因子。该方法将鸟枪法基因组表达文库的过表达以扰动细胞分裂过程，与高通量流式细胞分选技术相结合，用以筛选数以千计的克隆株。利用该策略，我们在模式生物大肠杆菌（Escherichia coli）中获得了呈现丝状形态的克隆株。遗传分析结果表明，本次筛选不仅鉴定出了已知的细胞分裂基因，还发现了此前未被报道参与细胞分裂过程的新基因。这种新型筛选策略可应用于包括病原菌在内的多种生物，其中细胞分裂相关基因及其调控因子是抗生素研发中极具潜力的药物靶点。