Department of Molecular Biology, Princeton University, Princeton, NJ, USA.
Rutgers Robert Wood Johnson Medical School, Piscataway, NJ, USA.
Methods Mol Biol. 2021;2357:125-146. doi: 10.1007/978-1-0716-1621-5_9.
Persisters are phenotypic variants within bacterial populations that tolerate antibiotic treatments considerably better than the majority of cells. A phenotypic quality that varies within bacterial populations is the chromosome number of individual cells. One, two, four, or more chromosomes per cell have been observed previously, and the impact of genome copy number can range from gene dosage effects to an inability to perform specific DNA repair functions, such as homologous recombination. We hypothesize that chromosome abundance is an underappreciated phenotypic variable that could impact persistence to antibiotics. Here, we describe methodologies to segregate bacterial populations based on chromosome number, assess the purity of those subpopulations, and suggest assays that could be used to quantify the impacts of genome abundance on persistence.
持留菌是细菌群体中的表型变异体，它们比大多数细胞更能耐受抗生素治疗。细菌群体中存在一种表型质量，即个别细胞的染色体数量。以前已经观察到每个细胞有一个、两个、四个或更多的染色体，基因组拷贝数的影响范围从基因剂量效应到无法执行特定的 DNA 修复功能，如同源重组。我们假设染色体丰度是一个未被充分认识的表型变量，可能会影响对抗生素的持久性。在这里，我们描述了基于染色体数量对细菌群体进行分类的方法，评估了这些亚群的纯度，并提出了可用于量化基因组丰度对持久性影响的测定方法。
