Flavin mononucleotide (FMN)-based fluorescent protein (FbFP) as reporter for gene expression in the anaerobe Bacteroides fragilis.

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  • Author(s): Lobo LA;Lobo LA; Smith CJ; Rocha ER
  • Source:
    FEMS microbiology letters [FEMS Microbiol Lett] 2011 Apr; Vol. 317 (1), pp. 67-74. Date of Electronic Publication: 2011 Feb 02.
  • Publication Type:
    Journal Article; Research Support, N.I.H., Extramural
  • Language:
    English
  • Additional Information
    • Source:
      Publisher: Oxford University Press Country of Publication: England NLM ID: 7705721 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1574-6968 (Electronic) Linking ISSN: 03781097 NLM ISO Abbreviation: FEMS Microbiol Lett Subsets: MEDLINE
    • Publication Information:
      Publication: 2015- : Oxford Oxford University Press
      Original Publication: Amsterdam, Published by Elsevier/North Holland on behalf of the Federation of European Microbiological Societies.
    • Subject Terms:
      Genes, Reporter*; Bacteroides fragilis/*genetics ; Coenzymes/*metabolism ; Flavin Mononucleotide/*metabolism ; Gene Expression Profiling/*methods ; Luminescent Proteins/*metabolism; Aerobiosis ; Anaerobiosis ; Animals ; Cell Line ; Fermentation ; Fluorescence ; Glucose/metabolism ; Macrophages/microbiology ; Maltose/metabolism ; Mice
    • Abstract:
      In this study, we show the expression of flavin mononucleotide-based fluorescent protein (FbFP) BS2 as a marker for gene expression in the opportunistic human anaerobic pathogen Bacteroides fragilis. Bacteroides fragilis 638R strain carrying osu∷bs2 constructs showed inducible fluorescence following addition of maltose anaerobically compared with nonfluorescent cells under glucose-repressed conditions. Bacteria carrying ahpC∷bs2 or dps∷bs2 constructs were fluorescent following induction by oxygen compared with nonfluorescent cells from the anaerobic control cultures. In addition, when these transcriptional fusion constructs were mobilized into B. fragilis IB263, a constitutive peroxide response strain, fluorescent BS2, was detected in both anaerobic and aerobic cultures, confirming the unique properties of the FbFP BS2 to yield fluorescent signal in B. fragilis in the presence and in the absence of oxygen. Moreover, intracellular expression of BS2 was also detected when cell culture monolayers of J774.1 macrophages were incubated with B. fragilis ahpC∷bs2 or dps∷bs2 strains within an anaerobic chamber. This suggests that ahpC and dps are induced following internalization by macrophages. Thus, we show that BS2 is a suitable tool for the detection of gene expression in obligate anaerobic bacteria in in vivo studies.
      (© 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.)
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    • Grant Information:
      R56 AI040588 United States AI NIAID NIH HHS; R21 AI079183 United States AI NIAID NIH HHS; R21 AI068659 United States AI NIAID NIH HHS; AI079183 United States AI NIAID NIH HHS; R21 AI079183-02 United States AI NIAID NIH HHS; R21 AI068659-02 United States AI NIAID NIH HHS; R01 AI040588 United States AI NIAID NIH HHS; AI068659 United States AI NIAID NIH HHS; AI40588 United States AI NIAID NIH HHS
    • Accession Number:
      0 (Coenzymes)
      0 (Luminescent Proteins)
      69-79-4 (Maltose)
      7N464URE7E (Flavin Mononucleotide)
      IY9XDZ35W2 (Glucose)
    • Publication Date:
      Date Created: 20110113 Date Completed: 20110608 Latest Revision: 20211020
    • Publication Date:
      20240829
    • Accession Number:
      PMC3053596
    • Accession Number:
      10.1111/j.1574-6968.2011.02212.x
    • Accession Number:
      21223361