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The essential genome of Streptococcus agalactiae.

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Publication Date: 2016-05-26

Journal Title: BMC Genomics

ISSN: 1471-2164

Volume: 17

Pages: 406

Language: English

Type: Article

This Version: VoR

Physical Medium: Electronic

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Citation: Hooven, T. A., Catomeris, A. J., Akabas, L. H., Randis, T. M., Maskell, D. J., Peters, S. E., Ott, S., et al. (2016). The essential genome of Streptococcus agalactiae

BMC Genomics, 17 406.

Abstract: BACKGROUND: Next-generation sequencing of transposon-genome junctions from a saturated bacterial mutant library (Tn-seq) is a powerful tool that permits genome-wide determination of the contribution of genes to fitness of the organism under a wide range of experimental conditions. We report development, testing, and results from a Tn-seq system for use in Streptococcus agalactiae (group B Streptococcus; GBS), an important cause of neonatal sepsis. METHODS: Our method uses a Himar1 mini-transposon that inserts at genomic TA dinucleotide sites, delivered to GBS on a temperature-sensitive plasmid that is subsequently cured from the bacterial population. In order to establish the GBS essential genome, we performed Tn-seq on DNA collected from three independent mutant libraries-with at least 135,000 mutants per library-at serial 24 h time points after outgrowth in rich media. RESULTS: After statistical analysis of transposon insertion density and distribution, we identified 13.5 % of genes as essential and 1.2 % as critical, with high levels of reproducibility. Essential and critical genes are enriched for fundamental cellular housekeeping functions, such as acyl-tRNA biosynthesis, nucleotide metabolism, and glycolysis. We further validated our system by comparing fitness assignments of homologous genes in GBS and a close bacterial relative, Streptococcus pyogenes, which demonstrated 93 % concordance. Finally, we used our fitness assignments to identify signal transduction pathway components predicted to be essential or critical in GBS. CONCLUSIONS: We believe that our baseline fitness assignments will be a valuable tool for GBS researchers and that our system has the potential to reveal key pathogenesis gene networks and potential therapeutic/preventative targets.


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Rights: Attribution 4.0 International

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Autor: Hooven, Thomas ACatomeris, Andrew JAkabas, Leor H Randis, Tara M Maskell, Duncan JohnPeters, Sarah EOtt, SandraSantana-Cruz, Ivet



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