Tn-seq: high-throughput parallel sequencing for fitness and genetic interaction studies in microorganisms Export

@article{citeulike:5910006, abstract = {Biological pathways are structured in complex networks of interacting genes. Solving the architecture of such networks may provide valuable information, such as how microorganisms cause disease. Here we present a method (Tn-seq) for accurately determining quantitative genetic interactions on a genome-wide scale in microorganisms. Tn-seq is based on the assembly of a saturated Mariner transposon insertion library. After library selection, changes in frequency of each insertion mutant are determined by sequencing the flanking regions en masse. These changes are used to calculate each mutant's fitness. Using this approach, we determined fitness for each gene of Streptococcus pneumoniae, a causative agent of pneumonia and meningitis. A genome-wide screen for genetic interactions of five query genes identified both alleviating and aggravating interactions that could be divided into seven distinct categories. Owing to the wide activity of the Mariner transposon, Tn-seq has the potential to contribute to the exploration of complex pathways across many different species.}, author = {van Opijnen, Tim and Bodi, Kip L. and Camilli, Andrew}, citeulike-article-id = {5910006}, citeulike-linkout-0 = {http://dx.doi.org/10.1038/nmeth.1377}, citeulike-linkout-1 = {http://dx.doi.org/10.1038/nmeth.1377}, day = {20}, doi = {10.1038/nmeth.1377}, issn = {1548-7091}, journal = {Nature Methods}, keywords = {e-map, genetic-interaction}, month = {September}, number = {10}, pages = {767--772}, posted-at = {2009-11-09 18:46:09}, priority = {2}, publisher = {Nature Publishing Group}, title = {Tn-seq: high-throughput parallel sequencing for fitness and genetic interaction studies in microorganisms}, url = {http://dx.doi.org/10.1038/nmeth.1377}, volume = {6}, year = {2009} }

TY - JOUR ID - citeulike:5910006 L3 - citeulike-article-id:5910006 N2 - Biological pathways are structured in complex networks of interacting genes. Solving the architecture of such networks may provide valuable information, such as how microorganisms cause disease. Here we present a method (Tn-seq) for accurately determining quantitative genetic interactions on a genome-wide scale in microorganisms. Tn-seq is based on the assembly of a saturated Mariner transposon insertion library. After library selection, changes in frequency of each insertion mutant are determined by sequencing the flanking regions en masse. These changes are used to calculate each mutant's fitness. Using this approach, we determined fitness for each gene of Streptococcus pneumoniae, a causative agent of pneumonia and meningitis. A genome-wide screen for genetic interactions of five query genes identified both alleviating and aggravating interactions that could be divided into seven distinct categories. Owing to the wide activity of the Mariner transposon, Tn-seq has the potential to contribute to the exploration of complex pathways across many different species. IS - 10 JF - Nature Methods SN - 1548-7091 EP - 772 TI - Tn-seq: high-throughput parallel sequencing for fitness and genetic interaction studies in microorganisms PB - Nature Publishing Group VL - 6 SP - 767 KW - e-map KW - genetic-interaction AU - van Opijnen, Tim AU - Bodi, Kip AU - Camilli, Andrew PY - 2009/09/20/ UR - http://dx.doi.org/10.1038/nmeth.1377 ER -