The DOI matches its Crossref record; 39 of the 39 references it cites were found in a registry or an index.
Tracking insertion mutants within libraries by deep sequencing and a genome-wide screen for Haemophilus genes required in the lung
Open accessAbstract
Rapid genome-wide identification of genes required for infection would expedite studies of bacterial pathogens. We developed genome-scale “negative selection” technology that combines high-density transposon mutagenesis and massively parallel sequencing of transposon/chromosome junctions in a mutant library to identify mutants lost from the library after exposure to a selective condition of interest. This approach was applied to comprehensively identify Haemophilus influenzae genes required to delay bacterial clearance in a murine pulmonary model. Mutations in 136 genes resulted in defects in vivo, and quantitative estimates of fitness generated by this technique were in agreement with independent validation experiments using individual mutant strains. Genes required in the lung included those with characterized functions in other models of H. influenzae pathogenesis and genes not previously implicated in infection. Genes implicated in vivo have reported or potential roles in survival during nutrient limitation, oxidative stress, and exposure to antimicrobial membrane perturbations, suggesting that these conditions are encountered by H. influenzae during pulmonary infection. The results demonstrate an efficient means to identify genes required for bacterial survival in experimental models of pathogenesis, and this approach should function similarly well in selections conducted in vitro and in vivo with any organism amenable to insertional mutagenesis.
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Cites
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Cited by
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Self-citationDeJesus, M. A., Gerrick, E. R., Xu, W., Park, S. W., Long, J. E., Boutte, C. C., Rubin, E. J., Schnappinger, D., Ehrt, S., Fortune, S. M., et al. (2017) · mBio · doi:10.1128/mbio.02133-16 · Cited by 668
Transposon insertion sequencing: a new tool for systems-level analysis of microorganisms
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Biofilms and Inflammation in Chronic Wounds
Zhao, G., Usui, M. L., Lippman, S. I., James, G. A., Stewart, P. S., Fleckman, P., & Olerud, J. E. (2013) · Advances in Wound Care · doi:10.1089/wound.2012.0381 · Cited by 448
Genome-wide identification of Acinetobacter baumannii genes necessary for persistence in the lung
Wang, N., Ozer, E. A., Mandel, M. J., & Hauser, A. R. (2014) · mBio · doi:10.1128/mbio.01163-14 · Cited by 234
Metadata
- Authors
- Gawronski, J. D., Wong, S. M. S., Giannoukos, G., Ward, D. V., & Akerley, B. J.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 2009-09-22
- Volume
- 106
- Issue
- 38
- Pages
- 16422-16427
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W2105546078
- Cited by
- 371
- References
- 39
- FWCI
- 8.20
- Type
- journal article
- Language
- en
- Open access
- green
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- Gawronski, J. D., Wong, S. M. S., Giannoukos, G., Ward, D. V., & Akerley, B. J. (2009). Tracking insertion mutants within libraries by deep sequencing and a genome-wide screen for Haemophilus genes required in the lung. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.0906627106
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