Electrotransformation of Listeria species

  • Janet E. Alexander
Chapter
Part of the Springer Lab Manuals book series (SLM)

Abstract

Electroporation offers an efficient system for the introduction of genetic material into listeriae. Conjugation and protoplast transformation of Listeria monocytogenes has been reported (Flamm et al. 1984, Vicente et al. 1987). However, transformation rates (∼ 103/ μg of DNA) produced by these methods are too low for use in many molecular techniques. To facilitate the entry of DNA by electroporation, sufficient intensity of current must be applied to produce the optimum number of permeablilized areas. The intensity of current is a function of the field strength and time constant used.

Keywords

Sucrose Magnesium Agar Penicillin Bacillus 

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References

  1. Bio-Rad Manual, (1994). Gene Pulser Transfection Apparatus Operating Instructions and Application Guide. Bio-Rad, Hercules, California, USA.Google Scholar
  2. Camilli, A., Portnoy, D. A., and Youngman, P. (1990). Insertional mutagenesis of Listeria monocytogenes with a novel Tn917 derivative that allows direct cloning of DNA flanking transposon insertions. Journal of Bacteriology. 172: 3738–3744.Google Scholar
  3. Flamm, R. K., Hinrichs, D. J., and Thomasow, M. (1984). Introduction of pAMB1 into Listeria monocytogenes by conjugation and homology between native Listeria monocytogenes plasmids. Infection and Immunity. 44: 157–161.Google Scholar
  4. Kok, J., Van Der Vossen, J. M. B. M., and Venema, G. (1984). Construction of plasmid cloning vectors for lactic streptococci which also replicate in Bacillus subtilis and Escherichia coli. Applied Environmental Microbiology. 48: 726–731.Google Scholar
  5. Miller, J. F. (1988). Bacterial Electroporation. Molecular Biology Reports. Bio-Rad Laboratories, Bio-Rad, Richmond, California, USA. 5: 1–4.Google Scholar
  6. Park, S. F., and Stewart, G. S. A. B. (1990). High-efficiency transformation of >Listeria monocytogenes by electroporation of penicillin-treated cells. Gene. 94: 129–132.CrossRefGoogle Scholar
  7. Shigekawa, K., and Dower, W. J. (1988). Electroporation of eukaryotes and prokaryotes: a general approach to the introduction of macromolecules into cells. Biotechniques. 6: 742–751.Google Scholar
  8. Trevors, J. T., Chassy, B. M., Dover, W. J., and Blaschnek, H. P. (1992). Electrotransformation of bacteria by Plasmid DNA. In; Guide to Electroporation and Electrofusion. Academic Press Inc, London.Google Scholar
  9. Vicente, M. F., Baquero, F., and Prez-D’az, J. C. 1987. A protoplast transformation system for Listeria spp. Plasmid. 18: 89–92.CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2000

Authors and Affiliations

  • Janet E. Alexander
    • 1
  1. 1.Department of Microbiology and Immunology, Medical Sciences BuildingUniversity of LeicesterLeicesterUK

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