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Volume 154, Issue 4

Identification of genes expressed during human infection Free

Abstract

To identify transcripts that are expressed during human infection, we used selective capture of transcribed sequences (SCOTS) with RNA isolated from pustules obtained from three volunteers infected with , and with RNA isolated from broth-grown bacteria used to infect volunteers. With SCOTS, competitive hybridization of tissue-derived and broth-derived sequences identifies genes that may be preferentially expressed . Among the three tissue specimens, we identified 531 genes expressed . Southern blot analysis of 60 genes from each tissue showed that 87 % of the identified genes hybridized better with cDNA derived from tissue specimens than with cDNA derived from broth-grown bacteria. RT-PCR on nine additional pustules confirmed expression of 10 of 11 selected genes in other volunteers. Of the 531 genes, 139 were identified in at least two volunteers. These 139 genes fell into several functional categories, including biosynthesis and metabolism, regulation, and cellular processes, such as transcription, translation, cell division, DNA replication and repair, and transport. Detection of genes involved in anaerobic and aerobic respiration indicated that likely encounters both microenvironments within the pustule. Other genes detected suggest an increase in DNA damage and stress . Genes involved in virulence in other bacterial pathogens and 32 genes encoding hypothetical proteins were identified, and may represent novel virulence factors. We identified three genes, , and , known to be required for virulence in humans. This is the first study to broadly define transcripts expressed by in humans.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): SCOTS, selective capture of transcribed sequences
SGM
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2008-04-01
2024-04-18
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Identification of Haemophilus ducreyi genes expressed during human infection
Microbiology 154, 1152 (2008); https://doi.org/10.1099/mic.0.2007/013953-0
/content/journal/micro/10.1099/mic.0.2007/013953-0
/content/journal/micro/10.1099/mic.0.2007/013953-0
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