Candidate Essential Genes in Burkholderia cenocepacia J2315 Identified by Genome-Wide TraDIS.
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.
Faculty of Science and Technology, School of Biosciences and Biotechnology, Universiti Kebangsaan Malaysia Bangi, Malaysia.
Yee-Chin
YC
Wong
Chemical and Life Sciences and Engineering Division, King Abdullah University of Science and TechnologyThuwal, Saudi Arabia; The Westmead Institute for Medical Research and The Marie Bashir Institute for Infectious Diseases and Biosecurity, The University of Sydney, SydneyNSW, Australia.
Moataz
M
Abd El Ghany
Chemical and Life Sciences and Engineering Division, King Abdullah University of Science and Technology Thuwal, Saudi Arabia.
Raeece
R
Naeem
Codon Genomics SB Selangor, Malaysia.
Kok-Wei
KW
Lee
Codon Genomics SB Selangor, Malaysia.
Yung-Chie
YC
Tan
Chemical and Life Sciences and Engineering Division, King Abdullah University of Science and Technology Thuwal, Saudi Arabia.
Arnab
A
Pain
Faculty of Science and Technology, School of Biosciences and Biotechnology, Universiti Kebangsaan Malaysia Bangi, Malaysia.
Sheila
S
Nathan
Burkholderia cenocepacia; TraDIS; essential genes; mutagenesis; transposon