ArticleGenome research2005
Construction of a mini-Tn5-luxCDABE mutant library in Pseudomonas aeruginosa PAO1: a tool for identifying differentially regulated genes.
Article in Genome research, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 89 papers.
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89 citing papers in PubMed, 166 citations in OpenAlex.
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- Application of a quantitative framework to improve the accuracy of a bacterial infection model.Proceedings of the National Academy of Sciences of the United States of America · 2023Article
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- A multiplexed, three-dimensional pooling and next-generation sequencing strategy for creating barcoded mutant arrays: construction of a Schizosaccharomyces pombe transposon insertion library.Nucleic acids research · 2022Article
- Putrescine and Its Metabolic Precursor Arginine Promote Biofilm and c-di-GMP Synthesis in Pseudomonas aeruginosa.Journal of bacteriology · 2022Article
- Construction and evaluation of a bioluminescentMicrobiology (Reading, England) · 2021Article
- Human organoid biofilm model for assessing antibiofilm activity of novel agents.NPJ biofilms and microbiomes · 2021Article
- Mapping the Efficacy and Mode of Action of Ethylzingerone [4-(3-Ethoxy-4-Hydroxyphenyl) Butan-2-One] as an Active Agent againstApplied and environmental microbiology · 2020Article
- The Phage-EncodedViruses · 2020Article
- Plant defensin antibacterial mode of action against Pseudomonas species.BMC microbiology · 2020Article
- Pseudomonas putida rDNA is a favored site for the expression of biosynthetic genes.Scientific reports · 2019Article
- Genomic and Phenotypic Diversity among Ten Laboratory Isolates ofJournal of bacteriology · 2019Article
- Efficient transposon mutagenesis mediated by an IPTG-controlled conditional suicide plasmid.BMC microbiology · 2018Article
- Secreted Phosphatase and Deoxyribonuclease Are Required by Pseudomonas aeruginosa To Defend against Neutrophil Extracellular Traps.Infection and immunity · 2018Article
- Identification and Characterization of Type IV Pili as the Cellular Receptor of Broad Host RangeViruses · 2018Article
- Replication of the Ordered, Nonredundant Library of Pseudomonas aeruginosa strain PA14 Transposon Insertion Mutants.Journal of visualized experiments : JoVE · 2018Article
- Genome-scale metabolic modeling of responses to polymyxins in Pseudomonas aeruginosa.GigaScience · 2018Article
29 more citing papers are in PubMed but not listed here.
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Authors and funding
7 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Pseudomonas aeruginosa is a major cause of nosocomial (hospital-derived) infections, is the predominant pathogen in chronic cystic fibrosis lung infections, and remains difficult to treat due to its high intrinsic antibiotic resistance. The completion of the P. aeruginosa PAO1 genome sequence provides the opportunity for genome-wide studies to increase our understanding of the pathogenesis and biology of this important pathogen. In this report, we describe the construction of a mini-Tn5-luxCDABE mutant library and a high-throughput inverse PCR method to amplify DNA flanking the site of insertion for sequencing and insertion site mapping. In addition to producing polar knockout mutations in nonessential genes, the promoterless luxCDABE reporter present in the transposon serves as a real-time reporter of gene expression for the inactivated gene. A total of 2519 transposon insertion sites were mapped, 77% of which were nonredundant insertions. Of the insertions within an ORF, -55% of total and unique insertion sites were transcriptional luxCDABE fusions. A bias toward low insertion-site density in the genome region that surrounds the predicted terminus of replication was observed. To demonstrate the utility of chromosomal lux fusions, we performed extensive regulatory screens to identify genes that were differentially regulated under magnesium or phosphate limitation. This approach led to the discovery of many known and novel genes necessary for these environmental adaptations, including genes involved in resistance to cationic antimicrobial peptides. This dual-purpose mutant library allows for functional and regulation studies and will serve as a resource for the research community to further our understanding of P. aeruginosa biology.
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