Candidate essential genes in Burkholderia cenocepacia J2315 identified by genome-wide TraDIS

Yee Chin Wong, Moataz Abd El Ghany, Raeece Naeem, Kok Wei Lee, Yung Chie Tan, Arnab Pain, Sheila Nathan

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Burkholderia cenocepacia infection often leads to fatal cepacia syndrome in cystic fibrosis patients. However, antibiotic therapy rarely results in complete eradication of the pathogen due to its intrinsic resistance to many clinically available antibiotics. Recent attention has turned to the identification of essential genes as the proteins encoded by these genes may serve as potential targets for development of novel antimicrobials. In this study, we utilized TraDIS (Transposon Directed Insertion-site Sequencing) as a genome-wide screening tool to facilitate the identification of B. cenocepacia genes essential for its growth and viability. A transposon mutant pool consisting of approximately 500,000 mutants was successfully constructed, with more than 400,000 unique transposon insertion sites identified by computational analysis of TraDIS datasets. The saturated library allowed for the identification of 383 genes that were predicted to be essential in B. cenocepacia. We extended the application of TraDIS to identify conditionally essential genes required for in vitro growth and revealed an additional repertoire of 439 genes to be crucial for B. cenocepacia growth under nutrient-depleted conditions. The library of B. cenocepacia mutants can subsequently be subjected to various biologically related conditions to facilitate the discovery of genes involved in niche adaptation as well as pathogenicity and virulence.

Original languageEnglish
Article number1288
JournalFrontiers in Microbiology
Volume7
Issue numberAUG
DOIs
Publication statusPublished - 22 Aug 2016

Fingerprint

Burkholderia cenocepacia
Essential Genes
Genome
Libraries
Virulence
Burkholderia Infections
Growth
Genes
Anti-Bacterial Agents
Genetic Association Studies
Cystic Fibrosis
Food

Keywords

  • Burkholderia cenocepacia
  • Essential genes
  • Mutagenesis
  • TraDIS
  • Transposon

ASJC Scopus subject areas

  • Microbiology
  • Microbiology (medical)

Cite this

Candidate essential genes in Burkholderia cenocepacia J2315 identified by genome-wide TraDIS. / Wong, Yee Chin; El Ghany, Moataz Abd; Naeem, Raeece; Lee, Kok Wei; Tan, Yung Chie; Pain, Arnab; Nathan, Sheila.

In: Frontiers in Microbiology, Vol. 7, No. AUG, 1288, 22.08.2016.

Research output: Contribution to journalArticle

Wong, Yee Chin ; El Ghany, Moataz Abd ; Naeem, Raeece ; Lee, Kok Wei ; Tan, Yung Chie ; Pain, Arnab ; Nathan, Sheila. / Candidate essential genes in Burkholderia cenocepacia J2315 identified by genome-wide TraDIS. In: Frontiers in Microbiology. 2016 ; Vol. 7, No. AUG.
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