Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection Export

@article{baba06, 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/).}, author = {Baba, Tomoya and Ara, Takeshi and Hasegawa, Miki and Takai, Yuki and Okumura, Yoshiko and Baba, Miki and Datsenko, Kirill A. and Tomita, Masaru and Wanner, Barry L. and Mori, Hirotada}, citeulike-article-id = {1570095}, doi = {doi:10.1038/msb4100050}, journal = {Molecular Systems Biology}, number = {1}, posted-at = {2007-08-16 21:26:54}, priority = {0}, title = {Construction of {{\em Escherichia coli}} {K-12} in-frame, single-gene knockout mutants: the {Keio} collection}, volume = {2}, year = {2006} }

TY - JOUR ID - baba06 L3 - citeulike-article-id:1570095 N2 - 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/). IS - 1 JF - Molecular Systems Biology TI - Construction of {{\em Escherichia coli}} {K-12} in-frame, single-gene knockout mutants: the {Keio} collection VL - 2 AU - Baba, Tomoya AU - Ara, Takeshi AU - Hasegawa, Miki AU - Takai, Yuki AU - Okumura, Yoshiko AU - Baba, Miki AU - Datsenko, Kirill AU - Tomita, Masaru AU - Wanner, Barry AU - Mori, Hirotada PY - 2006/// ER -