Construction of Escherichia coli K‐12 in‐frame, single‐gene knockout mutants: the Keio collection
Molecular Systems BiologyPublished 21 February 2006Open access
Tomoya Baba, Takeshi Ara, Miki Hasegawa, Yuki Takai, Yoshiko Okumura, Miki Baba
Citations8,234
SJR quartileQ1
SJR score4.78
SNIP1.72
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TL;DR
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.
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/).
Keywords
Biochemistry, Genetics and Molecular Biology
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MycorrhizaThe extraction and quantification of ergosterol from ectomycorrhizal fungi and roots
70 Citations1993Robert K. Antibus, Robert L. Sinsabaugh
Ergosterol was produced in all isolates examined, which represented the major orders of ectomycorrhizal fungi, and Calculations of viable fungal biomass associated with field-collected roots were in agreement with those reported by others using the method on resynthesized ectomy Corrhizae.
European Journal of BiochemistrySystematic search for zinc‐binding proteins in <i>Escherichia coli</i>
68 Citations2002Akira Katayama, Atsuko Tsujii +3 more
A systematic search for Escherichia coli proteins with the zinc-binding activity was performed using the assay of radioactive Zn(II) binding to total E. coli proteins fractionated by two methods of two-dimensional gel electrophoresis, and nine zinc- binding proteins were newly identified.
Australian Journal of EcologyLitter inputs to a tropical Australian rainforest stream
68 Citations1993Lee J. Benson, Richard G. Pearson
Freshwater BiologyComparison of leaf processing rates under different temperature regimes in three headwater streams
66 Citations1996Jacquelyn M. Rowe, Sean K. Meegan +3 more
Research in MicrobiologyFunctional genomics of in Japan
64 Citations2000Hirotada Mori, Katsumi Isono +2 more
A working project team in Japan has organized and begun construction of clones containing each of the putative open reading frames of E. coli, aiming at reaching complete understanding of this model organism in the near future.
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