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

@article{citeulike:2924246, 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. 10.1073/pnas.0511100103}, author = {Liberati, Nicole T. and Urbach, Jonathan M. and Miyata, Sachiko and Lee, Daniel G. and Drenkard, Eliana and Wu, Gang and Villanueva, Jacinto and Wei, Tao and Ausubel, Frederick M. }, citeulike-article-id = {2924246}, doi = {10.1073/pnas.0511100103}, journal = {Proceedings of the National Academy of Sciences}, keywords = {aeruginosa, mutant, pa14}, month = {February}, number = {8}, pages = {2833--2838}, posted-at = {2008-06-24 20:50:47}, priority = {2}, title = {An ordered, nonredundant library of Pseudomonas aeruginosa strain PA14 transposon insertion mutants}, url = {http://dx.doi.org/10.1073/pnas.0511100103}, volume = {103}, year = {2006} }

TY - JOUR ID - citeulike:2924246 L3 - citeulike-article-id:2924246 TI - An ordered, nonredundant library of Pseudomonas aeruginosa strain PA14 transposon insertion mutants JF - Proceedings of the National Academy of Sciences VL - 103 IS - 8 SP - 2833 EP - 2838 N2 - 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. 10.1073/pnas.0511100103 KW - aeruginosa KW - mutant KW - pa14 AU - Liberati, Nicole AU - Urbach, Jonathan AU - Miyata, Sachiko AU - Lee, Daniel AU - Drenkard, Eliana AU - Wu, Gang AU - Villanueva, Jacinto AU - Wei, Tao AU - Ausubel, Frederick PY - 2006/02/21/ UR - http://dx.doi.org/10.1073/pnas.0511100103 ER -