Identification of host-induced pathogen genes by differential fluorescence induction reporter systems.

Published

Journal Article

The ability to monitor a pathogen's gene expression program in response to the host environment is central to understanding host-microbe interactions. This protocol describes the application of a fluorescence-based promoter trap strategy, termed differential fluorescence induction (DFI), to identify and characterize bacterial genes that are preferentially expressed in infected tissues. In this approach, animals are infected with a library of bacteria expressing random GFP transcriptional gene fusions, and fluorescent bacteria are recovered directly from host tissues using fluorescence-activated cell sorting (FACS). This methodology allows for the identification of bacterial promoters induced in distinct anatomical sites and at different stages of infection. Furthermore, unlike other methodologies, the use of the GFP reporter allows for single cell, temporal and spatial monitoring of pathogen gene expression in infected animals. Library construction, promoter identification and analysis can be done in 4-8 weeks.

Full Text

Duke Authors

Cited Authors

  • Bumann, D; Valdivia, RH

Published Date

  • 2007

Published In

Volume / Issue

  • 2 / 4

Start / End Page

  • 770 - 777

PubMed ID

  • 17446876

Pubmed Central ID

  • 17446876

Electronic International Standard Serial Number (EISSN)

  • 1750-2799

Digital Object Identifier (DOI)

  • 10.1038/nprot.2007.78

Language

  • eng

Conference Location

  • England