Comprehensive Transposon Mutant Library of Pseudomonas aeruginosa
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 su...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 2003-11, Vol.100 (24), p.14339-14344 |
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| creator | Jacobs, Michael A. Alwood, Ashley Thaipisuttikul, Iyarit Spencer, David Haugen, Eric Ernst, Stephen Will, Oliver Kaul, Rajinder Raymond, Christopher Levy, Ruth Chun-Rong, Liu Guenthner, Donald Bovee, Donald Olson, Maynard V. Manoil, Colin |
| description | 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. |
| doi_str_mv | 10.1073/pnas.2036282100 |
| format | Article |
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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.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.2036282100</identifier><identifier>PMID: 14617778</identifier><language>eng</language><publisher>United States: National Academy of Sciences</publisher><subject>Biological Sciences ; Escherichia coli - genetics ; Essential genes ; Gene Library ; Genes ; Genes, Bacterial ; Genetic engineering ; Genetic transposition ; Genomes ; Libraries ; Library collections ; Microbiology ; Mutagenesis, Insertional ; Mutation ; Open reading frames ; Phenotype ; Phenotypes ; Pseudomonas aeruginosa ; Pseudomonas aeruginosa - genetics ; Species Specificity ; Transposons ; Twitching</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 2003-11, Vol.100 (24), p.14339-14344</ispartof><rights>Copyright 1993-2003 National Academy of Sciences of the United States of America</rights><rights>Copyright National Academy of Sciences Nov 25, 2003</rights><rights>Copyright © 2003, The National Academy of Sciences 2003</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c574t-3468e4a4ef82e600a24b9f52c87c83f96f97f3e29fee603a059d56b112476bf23</citedby><cites>FETCH-LOGICAL-c574t-3468e4a4ef82e600a24b9f52c87c83f96f97f3e29fee603a059d56b112476bf23</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/100/24.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/3148959$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/3148959$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,723,776,780,799,881,27901,27902,53766,53768,57992,58225</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/14617778$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Jacobs, Michael A.</creatorcontrib><creatorcontrib>Alwood, Ashley</creatorcontrib><creatorcontrib>Thaipisuttikul, Iyarit</creatorcontrib><creatorcontrib>Spencer, David</creatorcontrib><creatorcontrib>Haugen, Eric</creatorcontrib><creatorcontrib>Ernst, Stephen</creatorcontrib><creatorcontrib>Will, Oliver</creatorcontrib><creatorcontrib>Kaul, Rajinder</creatorcontrib><creatorcontrib>Raymond, Christopher</creatorcontrib><creatorcontrib>Levy, Ruth</creatorcontrib><creatorcontrib>Chun-Rong, Liu</creatorcontrib><creatorcontrib>Guenthner, Donald</creatorcontrib><creatorcontrib>Bovee, Donald</creatorcontrib><creatorcontrib>Olson, Maynard V.</creatorcontrib><creatorcontrib>Manoil, Colin</creatorcontrib><title>Comprehensive Transposon Mutant Library of Pseudomonas aeruginosa</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>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.</description><subject>Biological Sciences</subject><subject>Escherichia coli - genetics</subject><subject>Essential genes</subject><subject>Gene Library</subject><subject>Genes</subject><subject>Genes, Bacterial</subject><subject>Genetic engineering</subject><subject>Genetic transposition</subject><subject>Genomes</subject><subject>Libraries</subject><subject>Library collections</subject><subject>Microbiology</subject><subject>Mutagenesis, Insertional</subject><subject>Mutation</subject><subject>Open reading frames</subject><subject>Phenotype</subject><subject>Phenotypes</subject><subject>Pseudomonas aeruginosa</subject><subject>Pseudomonas aeruginosa - genetics</subject><subject>Species Specificity</subject><subject>Transposons</subject><subject>Twitching</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0T1v2zAQBmCiaNA4bucuRSF0CNBByfFDIjlkCIx-AQ7SIZ0JSj4mMiRSJaWg_felYSNuu2TicM97vMMR8pbCBQXJL0dv0wUDXjPFKMALsqCgaVkLDS_JAoDJUgkmTslZSlsA0JWCV-SUippKKdWCXK_CMEZ8QJ-6RyzuovVpDCn44maerJ-KdddEG38XwRXfE86bMIT8Z2ExzvedD8m-JifO9gnfHN4l-fH5093qa7m-_fJtdb0u20qKqeSiViisQKcY1gCWiUa7irVKtoo7XTstHUemHeYyt1DpTVU3lDIh68YxviRX-77j3Ay4adFP0fZmjN2Q5zPBdubfiu8ezH14NEzxSvOcPz_kY_g5Y5rM0KUW-956DHMyVLOayyyX5MN_cBvm6PNuhgHlQnBVZXS5R20MKUV0T4NQMLvTmN1pzPE0OfH-7_mP_nCLDD4ewC55bAeGiaw418bNfT_hrynb4hmbybs92aYpxCfDqVA6b_kHnH6s8A</recordid><startdate>20031125</startdate><enddate>20031125</enddate><creator>Jacobs, Michael A.</creator><creator>Alwood, Ashley</creator><creator>Thaipisuttikul, Iyarit</creator><creator>Spencer, David</creator><creator>Haugen, Eric</creator><creator>Ernst, Stephen</creator><creator>Will, Oliver</creator><creator>Kaul, Rajinder</creator><creator>Raymond, Christopher</creator><creator>Levy, Ruth</creator><creator>Chun-Rong, Liu</creator><creator>Guenthner, Donald</creator><creator>Bovee, Donald</creator><creator>Olson, Maynard V.</creator><creator>Manoil, Colin</creator><general>National Academy of Sciences</general><general>National Acad Sciences</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QG</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TK</scope><scope>7TM</scope><scope>7TO</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>5PM</scope></search><sort><creationdate>20031125</creationdate><title>Comprehensive Transposon Mutant Library of Pseudomonas aeruginosa</title><author>Jacobs, Michael A. ; 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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.</abstract><cop>United States</cop><pub>National Academy of Sciences</pub><pmid>14617778</pmid><doi>10.1073/pnas.2036282100</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| source | Jstor Complete Legacy; MEDLINE; PubMed Central; Alma/SFX Local Collection; Free Full-Text Journals in Chemistry |
| subjects | Biological Sciences Escherichia coli - genetics Essential genes Gene Library Genes Genes, Bacterial Genetic engineering Genetic transposition Genomes Libraries Library collections Microbiology Mutagenesis, Insertional Mutation Open reading frames Phenotype Phenotypes Pseudomonas aeruginosa Pseudomonas aeruginosa - genetics Species Specificity Transposons Twitching |
| title | Comprehensive Transposon Mutant Library of Pseudomonas aeruginosa |
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