Engineering Bacteriophage-Based Biosensors

  • Daniel Brownell
  • John King
  • Brian Caliando
  • Lada Sycheva
  • Michael KoerisEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1898)


Bacteriophages have been used for diagnostic purposes in the past, but a lack of parallelizable engineering methods had limited their applicability to a narrow subset of diagnostic settings. More recently, however, advances in DNA sequencing and the introduction of more sensitive reporter systems have enabled novel engineering methods, which in turn have broadened the scope of modern phage diagnostics. Here we describe advanced methods to engineer the genomes of bacteriophages in a modular and rapid fashion.

Key words

Bacteriophage engineering Genome engineering Reporter systems Diagnostics Luciferase 


  1. 1.
    Loessner MJ, Rees CE, Stewart GS, Scherer S (1996) Construction of luciferase reporter bacteriophage A511::luxAB for rapid and sensitive detection of viable listeria cells. Appl Environ Microbiol 62:1133–1140PubMedPubMedCentralGoogle Scholar
  2. 2.
    Klumpp J, Loessner MJ (2014) In: Thouand G, Marks R (eds) Bioluminescence: fundamentals and applications in biotechnology—volume 1. Springer, Berlin Heidelberg, p 155–171.
  3. 3.
    Loessner MJ, Rudolf M, Scherer S (1997) Evaluation of luciferase reporter bacteriophage A511::luxAB for detection of Listeria monocytogenes in contaminated foods. Appl Environ Microbiol 63:2961–2965PubMedPubMedCentralGoogle Scholar
  4. 4.
    Lu TK, Bowers J, Koeris MS (2013) Advancing bacteriophage-based microbial diagnostics with synthetic biology. Trends Biotechnol 31:325–327CrossRefGoogle Scholar
  5. 5.
    Karlin S, Burge C, Campbell AM (1992) Statistical analyses of counts and distributions of restriction sites in DNA sequences. Nucleic Acids Res 20:1363–1370CrossRefGoogle Scholar
  6. 6.
    Gelfand MS, Koonin EV (1997) Avoidance of palindromic words in bacterial and archaeal genomes: a close connection with restriction enzymes. Nucleic Acids Res 25:2430–2439CrossRefGoogle Scholar
  7. 7.
    Zhang H, Fouts DE, DePew J, Stevens RH (2013) Genetic modifications to temperate Enterococcus faecalis phage ϕEf11 that abolish the establishment of lysogeny and sensitivity to repressor, and increase host range and productivity of lytic infection. Microbiology 159:1023–1035CrossRefGoogle Scholar
  8. 8.
    Makowski L (1994) Phage display: structure, assembly and engineering of filamentous bacteriophage M13. Curr Opin Struct Biol 4:225–230CrossRefGoogle Scholar
  9. 9.
    Pouillot F, Blois H, Iris F (2010) Genetically engineered virulent phage banks in the detection and control of emergent pathogenic bacteria. Biosecurity Bioterrorism Biodefense Strategy Pract Sci 8:155–169CrossRefGoogle Scholar
  10. 10.
    Koeris MS, Shivers RP, Brownell, DR, Holder JW, Bowers JL (2014) Recombinant phage and bacterial detection methods.
  11. 11.
    Lu TKT et al (2013) Recombinant phage and methods.
  12. 12.
    Gibson DG et al (2009) Enzymatic assembly of DNA molecules up to several hundred kilobases. Nat Methods 6:343–345CrossRefGoogle Scholar
  13. 13.
    Klumpp J et al (2008) The terminally redundant, nonpermuted genome of Listeria bacteriophage A511: a model for the SPO1-like myoviruses of gram-positive bacteria. J Bacteriol 190:5753–5765CrossRefGoogle Scholar
  14. 14.
    Monk IR, Gahan CGM, Hill C (2008) Tools for functional postgenomic analysis of Listeria monocytogenes. Appl Environ Microbiol 74:3921–3934CrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  • Daniel Brownell
    • 1
  • John King
    • 1
  • Brian Caliando
    • 1
  • Lada Sycheva
    • 1
  • Michael Koeris
    • 1
    Email author
  1. 1.Sample 6 TechnologiesWoburnUSA

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