DOI: 10.1073/pnas.2036282100. Comprehensive transposon mutant library of <i>Pseudomonas aeruginosa</i>

Comprehensive transposon mutant library of Pseudomonas aeruginosa

10.1073/pnas.2036282100

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

We have developed technologies for creating saturating libraries of sequence-defined transposon insertion mutants in which each strain is maintained. Phenotypic analysis of such libraries should provide a virtually complete identification of nonessential genes required for any process for which a suitable screen can be devised. The approach was applied to Pseudomonas aeruginosa , an opportunistic pathogen with a 6.3-Mbp genome. The library that was generated consists of 30,100 sequence-defined mutants, corresponding to an average of five insertions per gene. About 12% of the predicted genes of this organism lacked insertions; many of these genes are likely to be essential for growth on rich media. Based on statistical analyses and bioinformatic comparison to known essential genes in E. coli , we estimate that the actual number of essential genes is 300-400. Screening the collection for strains defective in two defined multigenic processes (twitching motility and prototrophic growth) identified mutants corresponding to nearly all genes expected from earlier studies. Thus, phenotypic analysis of the collection may produce essentially complete lists of genes required for diverse biological activities. The transposons used to generate the mutant collection have added features that should facilitate downstream studies of gene expression, protein localization, epistasis, and chromosome engineering.

Bibliography

Jacobs, M. A., Alwood, A., Thaipisuttikul, I., Spencer, D., Haugen, E., Ernst, S., Will, O., Kaul, R., Raymond, C., Levy, R., Chun-Rong, L., Guenthner, D., Bovee, D., Olson, M. V., & Manoil, C. (2003). Comprehensive transposon mutant library of Pseudomonas aeruginosa. Proceedings of the National Academy of Sciences, 100(24), 14339â14344.

Authors 15

Michael A. Jacobs (first)
Ashley Alwood (additional)
Iyarit Thaipisuttikul (additional)
David Spencer (additional)
Eric Haugen (additional)
Stephen Ernst (additional)
Oliver Will (additional)
Rajinder Kaul (additional)
Christopher Raymond (additional)
Ruth Levy (additional)
Liu Chun-Rong (additional)
Donald Guenthner (additional)
Donald Bovee (additional)
Maynard V. Olson (additional)
Colin Manoil (additional)

References 15 Referenced 1,032

10.1038/nature00935
10.1073/pnas.0730515100
Berg C. M. & Berg D. E. (1995) in Mobile Genetic Elements ed. Sherratt D. (Oxford Univ. Press Oxford) pp. 38-68.
10.1016/S0076-6879(00)28418-8
10.1038/46558
10.1128/JB.184.16.4555-4572.2002
10.1111/j.1574-6968.2002.tb11162.x
10.1073/pnas.012602299
10.1101/gr.8.3.175
10.1038/nbt715
10.1038/35023079
Johnson N. L. & Kotz S. (1977) Urn Models and Their Application: An Approach to Modern Discrete Probability Theory (Wiley New York). (10.2307/2530628)
10.1099/00221287-145-10-2863
10.1146/annurev.micro.56.012302.160938
10.1038/nbt740

Dates

Type	When
Created	21 years, 9 months ago (Nov. 25, 2003, 3:59 p.m.)
Deposited	3 years, 4 months ago (April 25, 2022, 9:43 p.m.)
Indexed	2 weeks, 3 days ago (Aug. 19, 2025, 5:59 a.m.)
Issued	21 years, 9 months ago (Nov. 14, 2003)
Published	21 years, 9 months ago (Nov. 14, 2003)
Published Online	21 years, 9 months ago (Nov. 14, 2003)
Published Print	21 years, 9 months ago (Nov. 25, 2003)

Funders 0

None

BibTeX

@article{Jacobs_2003, title={Comprehensive transposon mutant library of Pseudomonas aeruginosa}, volume={100}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.2036282100}, DOI={10.1073/pnas.2036282100}, number={24}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Jacobs, Michael A. and Alwood, Ashley and Thaipisuttikul, Iyarit and Spencer, David and Haugen, Eric and Ernst, Stephen and Will, Oliver and Kaul, Rajinder and Raymond, Christopher and Levy, Ruth and Chun-Rong, Liu and Guenthner, Donald and Bovee, Donald and Olson, Maynard V. and Manoil, Colin}, year={2003}, month=nov, pages={14339–14344} }

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