Generation of Random luxCDABE Transcriptional Fusions in the Genome of Salmonella enterica

Methods Mol Biol. 2021:2182:141-151. doi: 10.1007/978-1-0716-0791-6_13.

Abstract

The luxCDABE operon of Photorhabdus luminescens can be used as a bioluminescent reporter to measure gene transcription nondestructively. Here we describe protocols to (1) generate random transcriptional fusions of the lux operon to genes of the Salmonella genome, (2) screen for specific fusions with constitutive expression, Salmonella pathogenicity island 1-related expression, or Salmonella pathogenicity island 2-related expression, and (3) determine the site of luxCDABE integration.

Keywords: Bioluminescence; Salmonella; Tn5; Type III secretion system; luxCDABE.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics*
  • Genes, Reporter / genetics
  • Genome, Bacterial / genetics*
  • Luminescent Measurements / methods
  • Operon / genetics
  • Photorhabdus / genetics*
  • Salmonella enterica / genetics*
  • Transcription, Genetic / genetics*

Substances

  • Bacterial Proteins