Live-Cell Visualization of DNA Transfer and Pilus Dynamics During Bacterial Conjugation

Kelly Goldlust; Agathe Couturier; Laurent Terradot; Christian Lesterlin

doi:10.1007/978-1-0716-2221-6_6

Live-Cell Visualization of DNA Transfer and Pilus Dynamics During Bacterial Conjugation

Methods Mol Biol. 2022:2476:63-74. doi: 10.1007/978-1-0716-2221-6_6.

Authors

Kelly Goldlust¹, Agathe Couturier¹, Laurent Terradot¹, Christian Lesterlin²

Affiliations

¹ Microbiologie Moléculaire et Biochimie Structurale (MMSB), Université Lyon 1, CNRS, Inserm, UMR5086, Lyon Cedex 07, France.
² Microbiologie Moléculaire et Biochimie Structurale (MMSB), Université Lyon 1, CNRS, Inserm, UMR5086, Lyon Cedex 07, France. christian.lesterlin@ibcp.fr.

PMID: 35635697
DOI: 10.1007/978-1-0716-2221-6_6

Abstract

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.

Keywords: Bacterial DNA conjugation; Conjugative pilus dynamics; DNA transfer dynamics; Horizontal gene transfer; Live-cell microscopy; Maleimide bioconjugation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Conjugation, Genetic*
DNA
Fimbriae, Bacterial* / genetics
Gene Transfer, Horizontal
Plasmids / genetics

Substances

DNA