Modulation of bft expression by the Bacteroides fragilis pathogenicity island and its flanking region

Augusto A. Franco, Rodney K. Cheng, Alan Goodman, Cynthia L. Sears

Research output: Contribution to journalArticlepeer-review

Abstract

To establish a recombinant system for high-level expression of biologically active Bacteroides fragilis toxin (BFT), we studied the expression of bft in non-toxigenic B. fragilis (NTBF) strains. The bft gene and the B. fragilis pathogenicity island (BfPAI) were cloned into NTBF strains with two distinct genetic patterns: (i) pattern II, strains lacking the BfPAI and its flanking region; and (ii) pattern III, strains lacking the BfPAI but containing its flanking region. Analysis of BFT activity of these recombinant strains on HT29/C1 cells showed that both the BfPAI and its flanking regions are important to optimal BFT activity. Reverse transcription polymerase chain reaction (RT-PCR) analysis indicated that the BfPAI and its flanking regions modulate bft expression. Further experiments demonstrated that the ≈ 700 bp region upstream of bft is the BfPAI region critical for optimal bft expression. We conclude that both the region flanking the BfPAI and ≈ 700 bp region upstream of bft are crucial to maximal BFT production by ETBF strains.

Original languageEnglish (US)
Pages (from-to)1067-1077
Number of pages11
JournalMolecular Microbiology
Volume45
Issue number4
DOIs
StatePublished - 2002

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

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