Microbial Transposon Mutagenesis pp 105-116 | Cite as
Transposon Mutagenesis of Bacteroides fragilis
Abstract
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 MutantsReferences
- 1.Sekirov I, Russell S, Antunes L (2010) Gut microbiota in health and disease. Physiol Rev 90:859–904. https://doi.org/10.1152/physrev.00045.2009 CrossRefPubMedGoogle Scholar
- 2.Round JL, Mazmanian SK (2009) The gut microbiota shapes intestinal immune responses during health and disease. Nat Rev Immunol 9:313–323. https://doi.org/10.1038/nri2515 CrossRefPubMedPubMedCentralGoogle Scholar
- 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. https://doi.org/10.1111/nyas.13660 CrossRefPubMedGoogle Scholar
- 4.Wexler HM (2007) Bacteroides: The good, the bad, and the nitty-gritty. Clin Microbiol Rev 20:593–621. https://doi.org/10.1128/CMR.00008-07 CrossRefPubMedPubMedCentralGoogle Scholar
- 5.Hecht DW (2004) Prevalence of antibiotic resistance in anaerobic bacteria: Worrisome developments. Clin Infect Dis 39:92–97. https://doi.org/10.1086/421558 CrossRefPubMedGoogle Scholar
- 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. https://doi.org/10.1128/AAC.00386-13 CrossRefPubMedPubMedCentralGoogle Scholar
- 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. https://doi.org/10.1186/1471-2164-15-429 CrossRefPubMedPubMedCentralGoogle Scholar
- 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. https://doi.org/10.1093/jac/dku219 CrossRefPubMedPubMedCentralGoogle Scholar
- 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. https://doi.org/10.1016/j.anaerobe.2018.02.005 CrossRefPubMedGoogle Scholar
- 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.Klein BA, Tenorio EL, Lazinski DW et al (2012) Identification of essential genes of the periodontal pathogen Porphyromonas gingivalis. BMC Genomics 13:578. https://doi.org/10.1186/1471-2164-13-578 CrossRefPubMedPubMedCentralGoogle Scholar
- 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.Veeranagouda Y, Husain F, Wexler HM (2013) Transposon mutagenesis of Bacteroides fragilis using a mariner transposon vector. Anaerobe 22:126–129. https://doi.org/10.1016/j.anaerobe.2013.04.012 CrossRefPubMedPubMedCentralGoogle Scholar