Measuring Plasmid Conjugation Using Fluorescent Reporters

Methods Mol Biol. 2020:2075:99-110. doi: 10.1007/978-1-4939-9877-7_7.

Abstract

Fluorescence-based methods are increasingly popular because they (1) offer a faster alternative to labor-intensive traditional methods, (2) enable the development of automated high-throughput screening procedures, and (3) allow direct visualization of biological processes. Here we describe three fluorescence-based methods applicable for the detection and quantitation of plasmid conjugation. The first method uses flow cytometry as a fast and reliable alternative to traditional plating methods. A second one employs fluorescence expression for high-throughput analysis of plasmid conjugation. Finally we review a third method that enables direct visualization of plasmid transfer under the microscope.

Keywords: Bacterial conjugation; Flow cytometry; Fluorescent proteins; Plasmid conjugation; SeqA.

MeSH terms

  • Bacteria / genetics*
  • Bacteria / metabolism
  • Conjugation, Genetic*
  • Flow Cytometry
  • Fluorescent Antibody Technique
  • Genes, Reporter*
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Microscopy, Fluorescence / methods
  • Optical Imaging / methods*
  • Plasmids / genetics*

Substances

  • Luminescent Proteins