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Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection. Mol. Syst. Biol 2:2006.0008 (2006)
| Citations: | 713 - 7 self |
BibTeX
@MISC{Baba06constructionof,
author = {Tomoya Baba and Takeshi Ara and Miki Hasegawa and Yuki Takai and Yoshiko Okumura and Miki Baba and Kirill A Datsenko and Masaru Tomita and Barry L Wanner and Hirotada Mori},
title = {Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection. Mol. Syst. Biol 2:2006.0008},
year = {2006}
}
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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
Keyphrases
keio collection escherichia coli k-12 in-frame single-gene knockout mutant flp recognition target site unknown gene function in-frame deletion kanamycin cassette single-gene deletion open-reading frame common strain background resistance cassette high-throughput study escherichia coli k-12 independent mutant chromosomal gene essential gene mutational effect one-step method genome-wide testing k-12 bw25113 unknown function gene regulatory network nonessential gene systematic analysis deleted gene new resource
