ArticleMicrobiologyOpen2013
Burkholderia cenocepacia conditional growth mutant library created by random promoter replacement of essential genes.
Article in MicrobiologyOpen, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed, 26 citations in OpenAlex.
- Optimized cumate toolkit for tunable protein expression duringApplied and environmental microbiology · 2026Article
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- Chemical genetic approaches for the discovery of bacterial cell wall inhibitors.RSC medicinal chemistry · 2023Review
- Gradients in gene essentiality reshape antibacterial research.FEMS microbiology reviews · 2022Review
- Methodological tools to study species of the genus Burkholderia.Applied microbiology and biotechnology · 2021Review
- LoxTnSeq: random transposon insertions combined with cre/lox recombination and counterselection to generate large random genome reductions.Microbial biotechnology · 2021Article
- Improved Dynamic Range of a Rhamnose-Inducible Promoter for Gene Expression inApplied and environmental microbiology · 2021Article
- Article
- The Effect of 2-Thiocyanatopyridine Derivative 11026103 onAntibiotics (Basel, Switzerland) · 2019Article
- Competitive Fitness of Essential Gene Knockdowns Reveals a Broad-Spectrum Antibacterial Inhibitor of the Cell Division Protein FtsZ.Antimicrobial agents and chemotherapy · 2018Article
- Essential Two-Component Systems Regulating Cell Envelope Functions: Opportunities for Novel Antibiotic Therapies.The Journal of membrane biology · 2018Review
- The Essential Genome of Burkholderia cenocepacia H111.Journal of bacteriology · 2017Article
- Comparative analysis of the Burkholderia cenocepacia K56-2 essential genome reveals cell envelope functions that are uniquely required for survival in species of the genus Burkholderia.Microbial genomics · 2017Article
- Evaluation of the electron transfer flavoprotein as an antibacterial target in Burkholderia cenocepacia.Canadian journal of microbiology · 2017Article
- Competitive Growth Enhances Conditional Growth Mutant Sensitivity to Antibiotics and Exposes a Two-Component System as an Emerging Antibacterial Target in Burkholderia cenocepacia.Antimicrobial agents and chemotherapy · 2017Article
- Candidate Essential Genes in Burkholderia cenocepacia J2315 Identified by Genome-Wide TraDIS.Frontiers in microbiology · 2016Article
- Layering genetic circuits to build a single cell, bacterial half adder.BMC biology · 2015Article
- Comparative metabolic systems analysis of pathogenic Burkholderia.Journal of bacteriology · 2014Article
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
Identification of essential genes by construction of conditional knockouts with inducible promoters allows the identification of essential genes and creation of conditional growth (CG) mutants that are then available as genetic tools for further studies. We used large-scale transposon delivery of the rhamnose-inducible promoter, PrhaB followed by robotic screening of rhamnose-dependent growth to construct a genomic library of 106 Burkholderia cenocepacia CG mutants. Transposon insertions were found where PrhaB was in the same orientation of widely conserved, well-characterized essential genes as well as genes with no previous records of essentiality in other microorganisms. Using previously reported global gene-expression analyses, we demonstrate that PrhaB can achieve the wide dynamic range of expression levels required for essential genes when the promoter is delivered randomly and mutants with rhamnose-dependent growth are selected. We also show specific detection of the target of an antibiotic, novobiocin, by enhanced sensitivity of the corresponding CG mutant (PrhaB controlling gyrB expression) within the library. Modulation of gene expression to achieve 30-60% of wild-type growth created conditions for specific hypersensitivity demonstrating the value of the CG mutant library for chemogenomic experiments. In summary, CG mutants can be obtained on a large scale by random delivery of a tightly regulated inducible promoter into the bacterial chromosome followed by a simple screening for the CG phenotype, without previous information on gene essentiality.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.