Nature Biotechnology
16, 851 - 856 (1998)
doi:10.1038/nbt0998-851
A genome-based approach for the identification of essential bacterial genesFabrizio Arigoni1, 2, Francois Talabot1, 2, Manuel Peitsch1, 3, Michael D. Edgerton1, 2, Eric Meldrum1, 4, Elisabeth Allet1, 2, Richard Fish1, 5, Therese Jamotte1, 2, Marie-Laure Curchod1, 2
& Hannes Loferer1, 6, *
1 Geneva Biomedical Research Institute, Glaxo Wellcome Research and Development S.A., Geneva, Switzerland.
2Current address: Serono Pharmaceutical Research Institute, Geneva, Switzerland.
3Current address: Glaxo Wellcome, Experimental Research S.A., Geneva, Switzerland.
4Current address: Glaxo Wellcome, Cellular Science, Stevenedge, UK.
5Current address: Biochimie Médicale, Centre Médical Universitaire, Geneva, Switzerland.
6Current address: Genome Pharmaceuticals Corporation, Munich, Germany.
*Corresponding author (e-mail: Hannes.Loferer@gpc-ag.com). 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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