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Global Transposon Mutagenesis and Essential Gene Analysis of Helicobacter pylori

Division of Human Biology, Fred Hutchinson Cancer Research Center, Seattle, Washington,¹ Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California²

Received 8 June 2004/ Accepted 23 August 2004

We have constructed a genome-saturating mutant library of the human gastric pathogen Helicobacter pylori. Microarray tracking of transposon mutants (MATT) allowed us to map the position of 5,363 transposon mutants in our library. While we generally found insertions well distributed throughout the genome, 344 genes had no detectable transposon insertions, and this list is predicted to be highly enriched for essential genes. Comparison to the essential gene set of other bacteria revealed a surprisingly limited overlap with all organisms tested (11%), while 55% were essential in some organisms but not others. We independently verified the essentiality of several gene products, including an HtrA family serine protease, a hypothetical protein with putative phospholipase D activity, and a riboflavin specific deaminase. A limited screen for motility mutants allowed us to estimate that 4.5% of the genome is dedicated to this virulence-associated phenotype.

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