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

doi:10.1046/j.1365-2958.2002.03077.x

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

School of Medicine

Research output: Contribution to journal › Article › peer-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 language	English (US)
Pages (from-to)	1067-1077
Number of pages	11
Journal	Molecular Microbiology
Volume	45
Issue number	4
DOIs	https://doi.org/10.1046/j.1365-2958.2002.03077.x
State	Published - 2002

ASJC Scopus subject areas

Microbiology
Molecular Biology

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title = "Modulation of bft expression by the Bacteroides fragilis pathogenicity island and its flanking region",

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.",

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doi = "10.1046/j.1365-2958.2002.03077.x",

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T1 - Modulation of bft expression by the Bacteroides fragilis pathogenicity island and its flanking region

AU - Franco, Augusto A.

AU - Cheng, Rodney K.

AU - Goodman, Alan

AU - Sears, Cynthia L.

PY - 2002

Y1 - 2002

N2 - 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.

AB - 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.

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

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