Tn-seq library, a tool to decrypt Salmonella gene function - INRAE - Institut national de recherche pour l’agriculture, l’alimentation et l’environnement Accéder directement au contenu
Poster De Conférence Année : 2022

Tn-seq library, a tool to decrypt Salmonella gene function

Résumé

Numerous genomes are available in databases and one of the biggest challenges is to understand the function of each gene. In bacteria, transposon mutant libraries have long been used to identify genes important for bacterial survival, growth or virulence. Recent advances in high throughput sequencing now allow the screening of millions of mutants in a single or few experiments using methods such as TraDIS or Tn-seq. These methods are sensitive enough to detect small defects in mutant fitness and allow to study genes but also intergenic regions, promoter regions and essential protein domains in coding sequences if the library is saturated with transposon insertions. Here, we present the construction and the characterization of a saturated library in Salmonella Typhimurium ATCC 14028 virulent strain, which is commonly used in research laboratories. This library was obtained using the Himar1 transposon that randomly inserts into TA dinucleotides sites onto the genome. Transposon mutant library was sequenced with an Illumina NextSeq 500. After read mapping, further analyses were carried out with the TRANSIT software. These analyses confirmed saturation of the transposon mutant library both on the chromosome and the virulence plasmid. This allowed us to determine the essential genes and those providing advantage or disadvantage for bacterial growth. This Tn-seq library is now used in several research projects and is available to the scientific community through collaboration

Domaines

Bactériologie
Fichier non déposé

Dates et versions

hal-03735722 , version 1 (21-07-2022)

Identifiants

  • HAL Id : hal-03735722 , version 1

Citer

Jérôme Trotereau, Romain Jouan, Delphine Naquin, Maxime Branger, Philippe Velge, et al.. Tn-seq library, a tool to decrypt Salmonella gene function. Journées d'Animation Scientifique de la FéRI 2022- FéRI Scientific Days 2022, Région Centre Val de Loire, Jul 2022, Tours, France. ⟨hal-03735722⟩
82 Consultations
0 Téléchargements

Partager

Gmail Facebook X LinkedIn More