Home LiteratureArticle Details
PMID: 16477005 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

An ordered, nonredundant library of Pseudomonas aeruginosa strain PA14 transposon insertion mutants.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 103 ·No. 8 ·2006-02-21 ·Pages 2833-8

Liberati NT, Urbach JM, Miyata S, Lee DG, Drenkard E, Wu G, Villanueva J, Wei T, Ausubel FM

Abstract

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.

MeSH Terms
DNA Transposable Elements/genetics Databases, Genetic Genes, Bacterial/genetics Genome, Bacterial Genomic Library Mutagenesis, Insertional/genetics Mutation Pseudomonas aeruginosa/genetics
Chemicals
DNA Transposable Elements
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Liberati Nicole T
Department of Genetics, Harvard Medical School, Boston, MA 02114, USA.
Urbach Jonathan M
Miyata Sachiko
Lee Daniel G
Drenkard Eliana
Wu Gang
Villanueva Jacinto
Wei Tao
Ausubel Frederick M
References (39)
39 references, click to expand
  1. Global transposon mutagenesis and a minimal Mycoplasma genome.
    Science. 1999 Dec 10;286(5447):2165-9 PMID: 10591650
  2. Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic pathogen.
    Nature. 2000 Aug 31;406(6799):959-64 PMID: 10984043
  3. Genomewide insertional mutagenesis in Streptomyces coelicolor reveals additional genes involved in morphological differentiation.
    Proc Natl Acad Sci U S A. 2000 Aug 15;97(17):9642-7 PMID: 10931952
  4. Genetic footprinting with mariner-based transposition in Pseudomonas aeruginosa.
    Proc Natl Acad Sci U S A. 2000 Aug 29;97(18):10191-6 PMID: 10963681
  5. Pseudomonas aeruginosa GacA, a factor in multihost virulence, is also essential for biofilm formation.
    Mol Microbiol. 2001 Jun;40(5):1215-26 PMID: 11401724
  6. Comprehensive identification of conditionally essential genes in mycobacteria.
    Proc Natl Acad Sci U S A. 2001 Oct 23;98(22):12712-7 PMID: 11606763
  7. Selection analyses of insertional mutants using subgenic-resolution arrays.
    Nat Biotechnol. 2001 Nov;19(11):1060-5 PMID: 11689852
  8. A genome-scale analysis for identification of genes required for growth or survival of Haemophilus influenzae.
    Proc Natl Acad Sci U S A. 2002 Jan 22;99(2):966-71 PMID: 11805338
  9. Identification of virulence genes in a pathogenic strain of Pseudomonas aeruginosa by representational difference analysis.
    J Bacteriol. 2002 Feb;184(4):952-61 PMID: 11807055
  10. Pseudomonas biofilm formation and antibiotic resistance are linked to phenotypic variation.
    Nature. 2002 Apr 18;416(6882):740-3 PMID: 11961556
  11. Functional profiling of the Saccharomyces cerevisiae genome.
    Nature. 2002 Jul 25;418(6896):387-91 PMID: 12140549
  12. Phosphorylation of the Pseudomonas aeruginosa response regulator AlgR is essential for type IV fimbria-mediated twitching motility.
    J Bacteriol. 2002 Aug;184(16):4544-54 PMID: 12142425
  13. Large-scale analysis of the meningococcus genome by gene disruption: resistance to complement-mediated lysis.
    Genome Res. 2003 Mar;13(3):391-8 PMID: 12618369
  14. The alternative sigma factor RpoN regulates the quorum sensing gene rhlI in Pseudomonas aeruginosa.
    FEMS Microbiol Lett. 2003 Mar 28;220(2):187-95 PMID: 12670680
  15. Essential Bacillus subtilis genes.
    Proc Natl Acad Sci U S A. 2003 Apr 15;100(8):4678-83 PMID: 12682299
  16. Use of the Galleria mellonella caterpillar as a model host to study the role of the type III secretion system in Pseudomonas aeruginosa pathogenesis.
    Infect Immun. 2003 May;71(5):2404-13 PMID: 12704110
  17. The Drosophila melanogaster toll pathway participates in resistance to infection by the gram-negative human pathogen Pseudomonas aeruginosa.
    Infect Immun. 2003 Jul;71(7):4059-66 PMID: 12819096
  18. Genome-wide insertional mutagenesis of Arabidopsis thaliana.
    Science. 2003 Aug 1;301(5633):653-7 PMID: 12893945
  19. Comprehensive transposon mutant library of Pseudomonas aeruginosa.
    Proc Natl Acad Sci U S A. 2003 Nov 25;100(24):14339-44 PMID: 14617778
  20. Genome-wide RNAi analysis of growth and viability in Drosophila cells.
    Science. 2004 Feb 6;303(5659):832-5 PMID: 14764878
  21. The broad host range pathogen Pseudomonas aeruginosa strain PA14 carries two pathogenicity islands harboring plant and animal virulence genes.
    Proc Natl Acad Sci U S A. 2004 Feb 24;101(8):2530-5 PMID: 14983043
  22. A large-scale RNAi screen in human cells identifies new components of the p53 pathway.
    Nature. 2004 Mar 25;428(6981):431-7 PMID: 15042092
  23. Genome-wide RNAi of C. elegans using the hypersensitive rrf-3 strain reveals novel gene functions.
    PLoS Biol. 2003 Oct;1(1):E12 PMID: 14551910
  24. SadB is required for the transition from reversible to irreversible attachment during biofilm formation by Pseudomonas aeruginosa PA14.
    J Bacteriol. 2004 Jul;186(14):4476-85 PMID: 15231779
  25. Systematic mutagenesis of the Escherichia coli genome.
    J Bacteriol. 2004 Aug;186(15):4921-30 PMID: 15262929
  26. Construction of an Enterococcus faecalis Tn917-mediated-gene-disruption library offers insight into Tn917 insertion patterns.
    J Bacteriol. 2004 Nov;186(21):7280-9 PMID: 15489440
  27. DNA amplification fingerprinting using arbitrary oligonucleotide primers.
    Appl Biochem Biotechnol. 1993 Aug-Sep;42(2-3):189-200 PMID: 7902694
  28. Common virulence factors for bacterial pathogenicity in plants and animals.
    Science. 1995 Jun 30;268(5219):1899-902 PMID: 7604262
  29. Genes and proteins of Escherichia coli K-12 (GenProtEC).
    Nucleic Acids Res. 1997 Jan 1;25(1):51-2 PMID: 9016503
  30. Use of model plant hosts to identify Pseudomonas aeruginosa virulence factors.
    Proc Natl Acad Sci U S A. 1997 Nov 25;94(24):13245-50 PMID: 9371831
  31. Factors affecting transposition of the Himar1 mariner transposon in vitro.
    Genetics. 1998 May;149(1):179-87 PMID: 9584095
  32. Initiation of biofilm formation in Pseudomonas fluorescens WCS365 proceeds via multiple, convergent signalling pathways: a genetic analysis.
    Mol Microbiol. 1998 May;28(3):449-61 PMID: 9632250
  33. Flagellar and twitching motility are necessary for Pseudomonas aeruginosa biofilm development.
    Mol Microbiol. 1998 Oct;30(2):295-304 PMID: 9791175
  34. Killing of Caenorhabditis elegans by Pseudomonas aeruginosa used to model mammalian bacterial pathogenesis.
    Proc Natl Acad Sci U S A. 1999 Jan 19;96(2):715-20 PMID: 9892699
  35. Molecular mechanisms of bacterial virulence elucidated using a Pseudomonas aeruginosa-Caenorhabditis elegans pathogenesis model.
    Cell. 1999 Jan 8;96(1):47-56 PMID: 9989496
  36. In vivo transposition of mariner-based elements in enteric bacteria and mycobacteria.
    Proc Natl Acad Sci U S A. 1999 Feb 16;96(4):1645-50 PMID: 9990078
  37. Global transposon mutagenesis and essential gene analysis of Helicobacter pylori.
    J Bacteriol. 2004 Dec;186(23):7926-35 PMID: 15547264
  38. A three-component regulatory system regulates biofilm maturation and type III secretion in Pseudomonas aeruginosa.
    J Bacteriol. 2005 Feb;187(4):1441-54 PMID: 15687209
  39. Positive correlation between virulence of Pseudomonas aeruginosa mutants in mice and insects.
    J Bacteriol. 2000 Jul;182(13):3843-5 PMID: 10851003
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.0511100103
Published
2006-02-21
Epub
2006-00-13
Pages
2833-8
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC1413827
Subset
IM
Grants
NIDDK NIH HHS · P30 DK040561 · United States
NIDDK NIH HHS · P30 DK040561-10 · United States
NHLBI NIH HHS · U01 HL066678 · United States
NHLBI NIH HHS · U01 HL66678 · United States
Corrections
ErratumIn
-
External Links
PubMed source DOI: 10.1073/pnas.0511100103 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2