Skip to main content

Electroporation of Legionella Species

  • Chapter
Electrotransformation of Bacteria

Part of the book series: Springer Lab Manuals ((SLM))

Abstract

The molecular analysis of Legionella pneumophila had been limited by the lack of methods to genetically manipulate the organism. Since bacteriophages and transduction have not been reported in L. pneumophila, methods of introducing DNA into the organism were limited to conjugation until 1990. Since then electroporation has rapidly replaced conjugation as the preferred method of introducing DNA into L. pneumophila for mutagenesis — mediated by transposons (Pope et al, 1994, Wiater et al, 1994, Pruckler et al, 1995, McClain and Engleberg, 1996, Hickey and Cianciotto, 1997) or by allelic exchange (Cianciotto and Fields, 1992, Sadosky et al, 1993, Abu Kwaik et al, 1997) and for complementation studies (Marra and Shuman, 1992, Berger et al, 1994, Abu Kwaik and Pederson, 1996).

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  • Abu Kwaik Y, Pederson LL (1996) The use of differential display-PCR to isolate and characterize a Legionella pneumophila locus induced during the intracellular infection of macrophages. Mol Microbiol 21 (3): 543–56.

    Article  CAS  Google Scholar 

  • Abu Kwaik Y, Gao LY, Harb OS, Stone BJ (1997) Transcriptional regulation of the macrophage-induced gene (gspA) of Legionella pneumophila and phenotypic characterization of a null mutant. Mol Microbiol 24 (3): 62942.

    Article  Google Scholar 

  • Berger KH, Isberg RR (1993) Two distinct defects in intracellular growth complemented by a single genetic locus in Legionella pneumophila. Mol Microbiol 7 (1): 7–19.

    Article  CAS  Google Scholar 

  • Berger KH, Merriam JJ, Isberg RR (1994) Altered intracellular targeting properties associated with mutations in the Legionella pneumophila dotA gene. Mol Microbiol 14 (4): 809–22.

    Article  CAS  Google Scholar 

  • Brand BC, Sadosky AB, Shuman HA (1994) The Legionella pneumophila icm locus: a set of genes required for intracellular multiplication in human macrophages. Mol Microbiol 14 (4): 797–808.

    Article  CAS  Google Scholar 

  • Cianciotto NP, Eisenstein BI, Engleberg NC, Shuman H (1989) Genetics and Molecular Pathogenesis of Legionella pneumophila an intracellular parasite of macrophages Mol Biol Med 6: 409–424.

    CAS  Google Scholar 

  • Cianciotto NP, Fields BS (1992) Legionella pneumophila mip gene potentiates intracellular infection of protozoa and human macrophages, Proc Natl Acad Sci USA, 89: 5188–5191.

    Google Scholar 

  • Hickey EK, Cianciotto NP (1997) An iron-and fur-repressed Legionella pneumophila gene that promotes intracellular infection and encodes a protein with similarity to the Escherichia coli aerobactin synthetases. Infect Immun 65 (1): 133–43.

    CAS  Google Scholar 

  • Marra A, Blander SJ, Horwitz MA, Shuman HA (1992) Identification of a Legionella pneumophila locus required for intracellular multiplication in human macrophages Proc Natl Acad Sci USA. 89: 9607–9611.

    Article  CAS  Google Scholar 

  • Marra A, Shuman HA (1992) Genetics of Legionella pneumophila virulence. Annu Rev Genet 26: 51–69.

    Article  CAS  Google Scholar 

  • McClain MS, Engleberg NC (1996) Construction of an alkaline phosphatase fusion-generating transposon, mTnlOphoA. Gene 170: 147–148.

    Article  CAS  Google Scholar 

  • Miller F (1994) Bacterial transformation by electroporation. Methods in Enzymology, vol 235. Academic Press, London, pp 375–385.

    Google Scholar 

  • O’Connell WA, Bangsborg JM, Cianciotto NP (1995) Characterization of a Legionella micdadei mip mutant. Infect and Immun 63 (8): 2840–2845.

    Google Scholar 

  • Pope CD, Dhand L, Cianciotto NP (1994) Random mutagenesis of Legionella pneumophila with mini-Tn10. FEMS Microbiol Lett 124 (1): 107–11.

    Article  CAS  Google Scholar 

  • Pruckler JM, Benson RF, Moyenuddin M, Martin WT, Fields BS (1995) Association of flagellum expression and intracellular growth of Legionella pneumophila. Infect Immun 63 (12) 4928–4932.

    CAS  Google Scholar 

  • Sadosky BA, Wiater LA, Shuman, HA (1993) Identification of Legionella pneumophila genes required for growth within and killing of human macrophages. Infect Immun 61 (12): 5361–5373.

    CAS  Google Scholar 

  • Sambrook J., Fritsch EF, Maniatis T. (1989) Molecular Cloning: A laboratory manual. Cold Spring Harbor Laboratory, New York, Cold Spring Harbor Laboratory Press.

    Google Scholar 

  • Wiater LA, Sadosky BA, Shuman (1994) Mutagenesis of Legionella pneumophila using Tn903dIIlacZ: identification of a growth-phase-regulated pigmentation gene. Mol Microbiol 11 (4): 641–653.

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2000 Springer-Verlag Berlin Heidelberg

About this chapter

Cite this chapter

Viswanathan, V.K., Cianciotto, N.P. (2000). Electroporation of Legionella Species. In: Eynard, N., Teissié, J. (eds) Electrotransformation of Bacteria. Springer Lab Manuals. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-662-04305-9_25

Download citation

  • DOI: https://doi.org/10.1007/978-3-662-04305-9_25

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-642-08593-2

  • Online ISBN: 978-3-662-04305-9

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics