Generation of Bioluminescent Group A Streptococcus for Biophotonic Imaging

Methods Mol Biol. 2020:2136:71-77. doi: 10.1007/978-1-0716-0467-0_6.

Abstract

Bioluminescence is a rapid and cost-saving technology that enables monitoring of bacteria in real time in animal infection models. Here, we report a method for labeling GAS with a set of plasmids we have developed and deposited with Addgene. These plasmids can be used to either integrate firefly luciferase (Ffluc) or red-shifted firefly luciferase (FflucRT) into the GAS genome or to introduce the reporters on plasmids which have been stabilized with a toxin-antitoxin system to prevent loss of plasmid in the absence of antibiotic selection.

Keywords: Bioluminescence; Biophotonic imaging; Firefly luciferase; Toxin–antitoxin system.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents
  • Diagnostic Imaging
  • Diagnostic Tests, Routine
  • Genes, Reporter / genetics
  • Genetic Engineering / methods*
  • Immunologic Tests
  • Luciferases, Firefly / genetics
  • Luminescent Measurements / methods*
  • Luminescent Proteins / biosynthesis
  • Luminescent Proteins / genetics*
  • Plasmids / genetics
  • Streptococcus pyogenes / genetics

Substances

  • Anti-Bacterial Agents
  • Luminescent Proteins
  • Luciferases, Firefly