{"doi":"10.1038/msb4100050","title":"Construction of\n                    <i>Escherichia coli</i>\n                    K‐12 in‐frame, single‐gene knockout mutants: the Keio collection","abstract":"We have systematically made a set of precisely defined, single-gene deletions of all nonessential genes in Escherichia coli K-12. Open-reading frame coding regions were replaced with a kanamycin cassette flanked by FLP recognition target sites by using a one-step method for inactivation of chromosomal genes and primers designed to create in-frame deletions upon excision of the resistance cassette. Of 4288 genes targeted, mutants were obtained for 3985. To alleviate problems encountered in high-throughput studies, two independent mutants were saved for every deleted gene. These mutants-the 'Keio collection'-provide a new resource not only for systematic analyses of unknown gene functions and gene regulatory networks but also for genome-wide testing of mutational effects in a common strain background, E. coli K-12 BW25113. We were unable to disrupt 303 genes, including 37 of unknown function, which are candidates for essential genes. Distribution is being handled via GenoBase (http://ecoli.aist-nara.ac.jp/).","journal":"Molecular Systems Biology","year":2006,"id":2384,"datarank":19.275919370741683,"base_score":8.999742789830492,"endowment":8.999742789830492,"self_citation_contribution":1.349961418474574,"citation_network_contribution":17.925957952267108,"self_endowment_contribution":1.349961418474574,"citer_contribution":17.925957952267108,"corpus_percentile":99.8,"corpus_rank":1366,"citation_count":8100,"citer_count":194,"citers_with_citation_signal":194,"citers_with_endowment":194,"datacite_reuse_total":0,"is_dataset":true,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2006-01-01","authors":[{"id":29258,"name":"Takeshi Ara","orcid":"0000-0003-1754-2837","position":1,"is_corresponding":false},{"id":29259,"name":"Miki Hasegawa","orcid":"0000-0003-0082-2576","position":2,"is_corresponding":false},{"id":29260,"name":"Yuki Takai","orcid":"0000-0001-6506-5313","position":3,"is_corresponding":false},{"id":29261,"name":"Yoshiko Okumura","orcid":null,"position":4,"is_corresponding":false},{"id":29262,"name":"Miki Baba","orcid":null,"position":5,"is_corresponding":false},{"id":29263,"name":"Kirill A. Datsenko","orcid":null,"position":6,"is_corresponding":false},{"id":29264,"name":"Masaru Tomita","orcid":"0000-0003-3423-377X","position":7,"is_corresponding":false},{"id":29265,"name":"Barry L. Wanner","orcid":"0000-0002-8703-1517","position":8,"is_corresponding":false},{"id":29266,"name":"Hirotada Mori","orcid":"0000-0003-3855-778X","position":9,"is_corresponding":false},{"id":29257,"name":"Tomoya Baba","orcid":"0000-0001-8986-5905","position":0,"is_corresponding":true}],"reference_count":53,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-03-01T18:20:47.508186Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}