TY - JOUR AU - Wong, Yee-Chin AU - Abd El Ghany, Moataz AU - Naeem, Raeece AU - Lee, Kok-Wei AU - Tan, Yung-Chie AU - Pain, Arnab AU - Nathan, Sheila PY - 2016 M3 - Original Research TI - Candidate Essential Genes in Burkholderia cenocepacia J2315 Identified by Genome-Wide TraDIS JO - Frontiers in Microbiology UR - https://www.frontiersin.org/articles/10.3389/fmicb.2016.01288 VL - 7 SN - 1664-302X N2 - 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. ER -