Transposon Mutagenesis of Bacteroides fragilis

  • Yaligara Veeranagouda
  • Fasahath Husain
  • Hannah M. Wexler
Part of the Methods in Molecular Biology book series (MIMB, volume 2016)


Bacteroides fragilis is Gram-negative obligatory anaerobe which usually resides in the gut of humans and animals. As an important member of the human gut microbiota it plays a vital role in digestion and absorption of nutrients as well as shaping of host immune system. B. fragilis is also infamous for causing serious infections. Treatment of B. fragilis infections caused emergence of multidrug-resistant strains. Molecular biology tools such as transposon mutagenesis help to decipher and understand commensal and pathogenic faces of B. fragilis. Using two mariner transposon vectors we describe the detailed methodology for the transposon mutagenesis of B. fragilis. We also describe two methods for the identification of transposon integration site (TIS) in transposon mutants. Transposon mutagenesis methods described in this chapter serve as a great tool for studying B. fragilis.

Key words

Transposon mutagenesis Bacteroides fragilis Mariner transposon Mutants 


  1. 1.
    Sekirov I, Russell S, Antunes L (2010) Gut microbiota in health and disease. Physiol Rev 90:859–904. CrossRefPubMedGoogle Scholar
  2. 2.
    Round JL, Mazmanian SK (2009) The gut microbiota shapes intestinal immune responses during health and disease. Nat Rev Immunol 9:313–323. CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Erturk-Hasdemir D, Kasper DL (2018) Finding a needle in a haystack: Bacteroides fragilis polysaccharide A as the archetypical symbiosis factor. Ann N Y Acad Sci 1417:116–129. CrossRefPubMedGoogle Scholar
  4. 4.
    Wexler HM (2007) Bacteroides: The good, the bad, and the nitty-gritty. Clin Microbiol Rev 20:593–621. CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Hecht DW (2004) Prevalence of antibiotic resistance in anaerobic bacteria: Worrisome developments. Clin Infect Dis 39:92–97. CrossRefPubMedGoogle Scholar
  6. 6.
    Husain F, Veeranagouda Y, Hsi J et al (2013) Two multidrug-resistant clinical isolates of bacteroides fragilis carry a novel metronidazole resistance nim gene (nimJ). Antimicrob Agents Chemother 57:3767–3774. CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Veeranagouda Y, Husain F, Tenorio EL, Wexler HM (2014) Identification of genes required for the survival of B. fragilis using massive parallel sequencing of a saturated transposon mutant library. BMC Genomics 15:429. CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Veeranagouda Y, Husain F, Boente R et al (2014) Deficiency of the ferrous iron transporter FeoAB is linked with metronidazole resistance in Bacteroides fragilis. J Antimicrob Chemother 69:2634–2643. CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Rashidan M, Azimirad M, Alebouyeh M et al (2018) Detection of B. fragilis group and diversity of bft enterotoxin and antibiotic resistance markers cepA, cfiA and nim among intestinal Bacteroides fragilis strains in patients with inflammatory bowel disease. Anaerobe 50:93–100. CrossRefPubMedGoogle Scholar
  10. 10.
    Goodman AL, McNulty NP, Zhao Y et al (2009) Identifying genetic determinants needed to establish a human gut symbiont in its habitat. Cell Host Microbe 6:279–289CrossRefGoogle Scholar
  11. 11.
    Klein BA, Tenorio EL, Lazinski DW et al (2012) Identification of essential genes of the periodontal pathogen Porphyromonas gingivalis. BMC Genomics 13:578. CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Veeranagouda Y, Husain F, Wexler HM (2012) Transposon mutagenesis of the anaerobic commensal, Bacteroides fragilis, using the EZ::TN5 transposome. FEMS Microbiol Lett 333:94–100CrossRefGoogle Scholar
  13. 13.
    Veeranagouda Y, Husain F, Wexler HM (2013) Transposon mutagenesis of Bacteroides fragilis using a mariner transposon vector. Anaerobe 22:126–129. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Yaligara Veeranagouda
    • 1
    • 2
  • Fasahath Husain
    • 1
  • Hannah M. Wexler
    • 1
    • 2
  1. 1.Greater Los Angeles VA Health Care SystemLos AngelesUSA
  2. 2.David Geffen School of MedicineUniversity of California Los AngelesLos AngelesUSA

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