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A genome-based approach for the identification of essential bacterial genes

Nature Biotechnology volume 16, pages 851–856 (1998)Cite this article

Abstract

We have used comparative genomics to identify 26 Escherichia coli open reading frames that are both of unknown function (hypothetical open reading frames or y-genes) and conserved in the compact genome of Mycoplasma genitalium. Not surprisingly, these genes are broadly conserved in the bacterial world. We used a markerless knockout strategy to screen for essential E. coli genes. To verify this pheno-type, we constructed conditional mutants in genes for which no null mutants could be obtained. In total we identified six genes that are essential for E. coli (yhbZ, ygjD, ycfB, yfil, yihA, and yjeQ). The respective orthologs of the genes yhbZ, ygjD, ycfB, yjeQ, and yihA are also essential in Bacillus subtilis. This low number of essential genes was unexpected and might be due to a characteristic of the versatile genomes of E. coli and B subtilis that is comparable to the phenomenon of nonorthologous gene displacement. The gene ygjD, encoding a sialoglycoprotease, was eliminated from a minimal genome computationally derived from a comparison of the Haemophilus influenzae and M. genitalium genomes. We show that ygjD and its ortholog ydiE are essential in E. coli and B. subtilis, respectively. Thus, we include this gene in a minimal genome. This study systematically integrates comparative genomics and targeted gene disruptions to identify broadly conserved bacterial genes of unknown function required for survival on complex media.

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Author notes

  1. Fabrizio Arigoni, Francois Talabot, Michael D. Edgerton, Elisabeth Allet, Therese Jamotte & Marie-Laure Curchod

    Present address: Serono Pharmaceutical Research Institute, Geneva, Switzerland

  2. Manuel Peitsch

    Present address: Glaxo Wellcome, Experimental Research S.A., Geneva, Switzerland

  3. Eric Meldrum

    Present address: Glaxo Wellcome, Cellular Science, Stevenedge, UK

  4. Richard Fish

    Present address: Biochimie Médicale, Centre Médical Universitaire, Geneva, Switzerland

  5. Hannes Loferer

    Present address: Genome Pharmaceuticals Corporation, Munich, Germany

  6. Hannes Loferer: Corresponding author (e-mail: Hannes.Loferer@gpc-ag.com).

Authors and Affiliations

  1. Geneva Biomedical Research Institute, Glaxo Wellcome Research and Development S.A., Geneva, Switzerland

    Fabrizio Arigoni, Francois Talabot, Manuel Peitsch, Michael D. Edgerton, Eric Meldrum, Elisabeth Allet, Richard Fish, Therese Jamotte, Marie-Laure Curchod & Hannes Loferer

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Arigoni, F., Talabot, F., Peitsch, M. et al. A genome-based approach for the identification of essential bacterial genes. Nat Biotechnol 16, 851–856 (1998). https://doi.org/10.1038/nbt0998-851

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