PLoS Genetics (Jul 2009)

A strand-specific RNA-Seq analysis of the transcriptome of the typhoid bacillus Salmonella typhi.

  • Timothy T Perkins,
  • Robert A Kingsley,
  • Maria C Fookes,
  • Paul P Gardner,
  • Keith D James,
  • Lu Yu,
  • Samuel A Assefa,
  • Miao He,
  • Nicholas J Croucher,
  • Derek J Pickard,
  • Duncan J Maskell,
  • Julian Parkhill,
  • Jyoti Choudhary,
  • Nicholas R Thomson,
  • Gordon Dougan

DOI
https://doi.org/10.1371/journal.pgen.1000569
Journal volume & issue
Vol. 5, no. 7
p. e1000569

Abstract

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High-density, strand-specific cDNA sequencing (ssRNA-seq) was used to analyze the transcriptome of Salmonella enterica serovar Typhi (S. Typhi). By mapping sequence data to the entire S. Typhi genome, we analyzed the transcriptome in a strand-specific manner and further defined transcribed regions encoded within prophages, pseudogenes, previously un-annotated, and 3'- or 5'-untranslated regions (UTR). An additional 40 novel candidate non-coding RNAs were identified beyond those previously annotated. Proteomic analysis was combined with transcriptome data to confirm and refine the annotation of a number of hpothetical genes. ssRNA-seq was also combined with microarray and proteome analysis to further define the S. Typhi OmpR regulon and identify novel OmpR regulated transcripts. Thus, ssRNA-seq provides a novel and powerful approach to the characterization of the bacterial transcriptome.