Correction for Liberati et al., An ordered, nonredundant library of Pseudomonas aeruginosa strain PA14 transposon insertion mutants

By Nicole T. Liberati, Jonathan M. Urbach, S. Miyata, Daniel G. Lee, E. Drenkard, Gang Wu, J. Villanueva, Tao Wei, Frederick M. Ausubel. Proceedings of the National Academy of Sciences of the United States of America, 2006

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Random transposon insertion libraries have proven invaluable in studying bacterial genomes. Libraries that approach saturation must be large, with multiple insertions per gene, making comprehensive genome-wide scanning difficult. To facilitate genome-scale study of the opportunistic human pathogen Pseudomonas aeruginosa strain PA14, we constructed a nonredundant library of PA14 transposon mutants (the PA14NR Set) in which nonessential PA14 genes are represented by a single transposon insertion chosen from a comprehensive library of insertion mutants. The parental library of PA14 transposon insertion mutants was generated by using MAR2xT7, a transposon compatible with transposon-site hybridization and based on mariner. The transposon-site hybridization genetic footprinting feature broadens the utility of the library by allowing pooled MAR2xT7 mutants to be individually tracked under different experimental conditions. A public, internet-accessible database (the PA14 Transposon Insertion Mutant Database, http://ausubellab.mgh.harvard.edu/cgi-bin/pa14/home.cgi) was developed to facilitate construction, distribution, and use of the PA14NR Set. The usefulness of the PA14NR Set in genome-wide scanning for phenotypic mutants was validated in a screen for attachment to abiotic surfaces. Comparison of the genes disrupted in the PA14 transposon insertion library with an independently constructed insertion library in P. aeruginosa strain PAO1 provides an estimate of the number of P. aeruginosa essential genes.

Subfields: Genetics, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/01c63dd6582a975b86a9a4d763268783bdfd9e09

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Citations in the corpus

Genomic analysis reveals that Pseudomonas aeruginosa virulence is combinatorial (2006, methodology): “To screen for avirulent PA14 mutants, we utilized a nonredundant transposon insertion library [25].”
Simultaneous assay of every Salmonella Typhi gene using one million transposon mutants. (2009, neutral): “sertions per gene (Salama et al. 2004; Glass et al. 2006; Liberati et al. 2006; Gallagher et al. 2007; Laia et al. 2009).”
Pseudomonas genomes: diverse and adaptable. (2011, background): “Similarly, 20 530 unique transposon insertions were mapped within the PA14 genome (Liberati et al., 2006).”
DEG 5.0, a database of essential genes in both prokaryotes and eukaryotes (2008, background): “…subtilis (24), those determined by global transposon mutagenesis in Francisella novicida (25), Helicobacter pylori (26), Mycobacterium tuberculosis (27), Mycoplasma pulmonis (28) and Pseudomonas aeruginosa (29,30), and those determined by trapping lethal insertions in Salmonella typhimurium (31).”
A quorum-sensing inhibitor blocks Pseudomonas aeruginosa virulence and biofilm formation (2013, neutral): “aeruginosa PA14 ordered transposon library (31).”
DEG 10, an update of the database of essential genes that includes both protein-coding genes and noncoding genomic elements (2013, methodology): “Porphyromonasgingivalis 463 Tn-seq Yes (42) Pseudomonas aeruginosa 335 Transposon mutagenesis followed by genetic footprinting Yes (61)”
Genome-scale models of bacterial metabolism: reconstruction and applications (2008, background): “In parallel to this, collections of knockout mutants are being built for a growing number of bacteria (Akerley et al ., 2002; Jacobs et al ., 2003; Kobayashi et al ., 2003; Baba et al ., 2006; Liberati et al ., 2006; Suzuki et al ., 2006; de Berardinis et al ., 2008).”
Pseudomonas Genome Database: improved comparative analysis and population genomics capability for Pseudomonas genomes (2010, background): “Annotation updates are also made to other Pseudomonas genomes in the database through the process of contacting curators at other genome centers including those responsible for the ongoing annotation of the P. syringae DC3000 (35), P. aeruginosa PA14 (2) and P. aeruginosa LESB58 (36) reference strai”