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Electroporation Protocols for Microorganisms pp 115–123Cite as

Electrotransformation in Salmonella

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 47))

Abstract

Enteric bacteria are not naturally competent for DNA-mediated transformation, but methods derived from the Ca2+-shock method of Mandel and Higa (1) permit uptake of DNA at levels adequate for molecular cloning. Transformation frequencies using Ca2+-shock methods with selected strains of E. coli K-12 range from 106 to 108 transformants/µg of covalently closed circular plasmid DNA, but efficiency is frequently lower because these methods are very sensitive to purity of reagents, cleanliness and quality of glassware and plasticware, and methods of cell handling (2).

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References

  1. Mandel, M. and Higa, A. (1970) Calcium dependent bacteriophage DNA infection. J. Mol. Biol. 53, 157–162.

    Article  Google Scholar 

  2. Hanahan, D. (1987) in Escherichia coli and Salmonella typhimurium: Cellular and Molecular Biology (Low, K. B., Magasanik, B., Schaechter, M., and Umbarger, H. E., eds.), American Society for Microbiology, Washington, DC, pp. 1177–1183.

    Google Scholar 

  3. Lederberg, E. M. and Cohen, S. N. (1974) Transformation of Salmonella typhimurium by plasmid deoxyribonucleic acid. J. Bacteriol. 119, 1072–1074.

    PubMed  CAS  Google Scholar 

  4. Bursztyn, H., Sgaramella, V., Ciferri, O., and Lederberg, J. (1975) Transfectability of rough strains of Salmonella typhimurium. J. Bacteriol. 124, 1630–1634.

    PubMed  CAS  Google Scholar 

  5. MacLachlan, P. R. and Sanderson, K. E. (1985) Transformation of Salmonella typhimurium with plasmid DNA: differences between rough and smooth strains. J. Bacteriol. 161, 442–445.

    PubMed  CAS  Google Scholar 

  6. Dower, W. J., Miller, J. F., and Ragsdale, C. W. (1988) High efficiency transformation of Escherichia coli by high voltage electroporation. Nucleic Acids Res. 16, 6127–6145.

    Article  PubMed  CAS  Google Scholar 

  7. Shigekawa, K. and Dower, W. (1988) Electroporation of eukaryotes and prokaryotes: a general approach to the introduction of macromolecules into cells. Biotechniques 6, 742–750.

    PubMed  CAS  Google Scholar 

  8. MacLachlan, P. R., Kadam, S. K., and Sanderson, K. E. (1991) Cloning, characterization, and DNA sequence of the rfaLK region for lipopolysaccharide synthesis in Salmonella typhimurium LT2. J. Bacteriol. 173, 7151–7163.

    PubMed  CAS  Google Scholar 

  9. Sirisena, D. M., Brozek, K. A., MacLachlan, P. R., Sanderson, K. E., and Raetz, C. R. H. (1992) The rfaC gene of Salmonella typhimurium. J. Biol. Chem. 267, 18,874–18,884.

    PubMed  CAS  Google Scholar 

  10. Roantree, R. J., Kuo, T.-T., and MacPhee, D. G. (1977) The effect of defined polysaccharide core defects upon antibiotic resistances of Salmonella typhimurium. J. Gen. Microbiol. 103, 223–234.

    PubMed  CAS  Google Scholar 

  11. Wilkinson, R. G., Gemski, P., Jr., and Stocker, B. A. D. (1972) Non-smooth mutants of Salmonella typhimurium: differentiation by phage sensitivity and genetic mapping. J. Gen. Microbiol. 70, 527–554.

    PubMed  CAS  Google Scholar 

  12. Binotto, J., MacLachlan, P. R., and Sanderson, K. E. (1991) Electrotransformation in Salmonella typhimurium LT2. Can. J. Microbiol. 37, 474–477.

    Article  PubMed  CAS  Google Scholar 

  13. Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.

    Google Scholar 

  14. Powell, I. B., Acken, M. G., Hillier, A. J., and Davidson, B. E. (1988) A simple and rapid method for genetic transformation of lactic streptococci by electroporation. Appl. Environ. Microbiol. 54, 655–660.

    PubMed  CAS  Google Scholar 

  15. Bullas, L. R. and Ryu, J.-I (1983) Salmonella typhimurium LT2 strains which are r-m+ for all three chromosomally located systems of DNA restriction and modification. J. Bacteriol. 156, 471–474.

    PubMed  CAS  Google Scholar 

  16. Sanderson, K. E., Janzer, J., and Head, J. (1981) Influence of lipopolysaccharide and protein in the cell envelope on recipient capacity in conjugation of Salmonella typhimurium. J. Bacteriol. 148, 283–293.

    PubMed  CAS  Google Scholar 

  17. Schnaitman, C. A., Parker, C. T., Klena, J. D., Pradel, E. L., Pearson, N. B., Sanderson, K. E., and MacLachlan, P. R. (1991) Physical maps of the rfa loci of Escherichia coli K-12 and Salmonella typhimurium. J. Bacteriol. 173, 7410,7411.

    PubMed  CAS  Google Scholar 

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Author information

Author notes

  1. P. Ronald MacLachlan

    Present address: Veterinary Infectious Disease Organization, University of Saskatchewan, Saskatoon, Canada

Authors and Affiliations

  1. Salmonella Genetic Stock Centre, Department of Biological Sciences, University of Calgary, Canada

    Kenneth E. Sanderson, P. Ronald MacLachlan & Andrew Hessel

Authors
  1. Kenneth E. Sanderson
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  2. P. Ronald MacLachlan
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  3. Andrew Hessel
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Editor information

Editors and Affiliations

  1. Department of Cancer Biology, Harvard University School of Public Health, Boston, MA

    Jac A. Nickoloff

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© 1995 Humana Press Inc.

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Cite this protocol

Sanderson, K.E., MacLachlan, P.R., Hessel, A. (1995). Electrotransformation in Salmonella . In: Nickoloff, J.A. (eds) Electroporation Protocols for Microorganisms. Methods in Molecular Biology™, vol 47. Humana Press. https://doi.org/10.1385/0-89603-310-4:115

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  • DOI: https://doi.org/10.1385/0-89603-310-4:115

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-310-8

  • Online ISBN: 978-1-59259-534-1

  • eBook Packages: Springer Protocols

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