Microbiologie Moléculaire et Biochimie Structurale (MMSB), Université Lyon 1, CNRS, Inserm, UMR5086, Lyon Cedex 07, France.
Methods Mol Biol. 2022;2476:63-74. doi: 10.1007/978-1-0716-2221-6_6.
Bacterial genomes are highly plastic and evolve rapidly by acquiring new genetic information through horizontal gene transfer mechanisms. Capturing DNA transfer by conjugation between bacterial cells in real time is relevant to address bacterial genomes' dynamic architecture comprehensively. Here, we describe a method allowing the direct visualization of bacterial conjugation in live cells, including the fluorescent labeling of the conjugative pilus and the monitoring of plasmid DNA transfer from donor to recipient cells.
细菌基因组具有高度的可塑性,并通过水平基因转移机制获得新的遗传信息而快速进化。实时捕获细菌细胞之间的接合过程中的 DNA 转移对于全面了解细菌基因组的动态结构至关重要。在这里,我们描述了一种允许在活细胞中直接可视化细菌接合的方法,包括对接合性菌毛进行荧光标记以及监测质粒 DNA 从供体向受体细胞的转移。