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Production of cDNA Libraries by Electroporation

  • Protocol
Electroporation Protocols for Microorganisms

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

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Abstract

In the late 1970s and early 1980s, several laboratories (1–4) reported that an applied electric field could induce transient pores in eukaryotic cell membranes. In addition to small molecules, such as sucrose and dyes, DNA could be introduced into these electropermeabilized cells (5–7). More recently, the electroporation process has been used to transform bacterial cells with plasmid DNA efficiently (8,9).

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References

  1. Kinosita, K., Jr. and Tsong, T. Y. (1977) Hemolysis of human erythrocytes by a transient electric field. Proc. Natl. Acad. Sci. USA 74, 1923–1927.

    PubMed  CAS  Google Scholar 

  2. Baker, P. F. and Knight, D. E. (1978) A high voltage technique for gaining rapid access to the interior of secretory cells. J. Physiol. 284, 30.

    Google Scholar 

  3. Gauger, B. and Bentrup, F. W. (1979) A study of dielectric membrane breakdown in the Fucus egg. J. Membrane Biol. 48, 249–264.

    CAS  Google Scholar 

  4. Zimmermann, U., Vienken, J., and Pilwat, G. (1980) Development of drug carrier systems: electric field induced effects in cell membranes. J. Electroanal. Chem. 116, 553–574.

    Google Scholar 

  5. Wong, T. K. and Neumann, E. (1982) Electric field mediated gene transfer. Biochem. Biophys. Res. Commun. 107, 584–587.

    PubMed  CAS  Google Scholar 

  6. Potter, H., Weir, L., and Leder, P. (1984) Enhancer-dependent expression of human k immunoglobulin genes introduced into mouse pre-B lymphocytes by electroporation. Proc. Natl Acad. Sci. USA 81, 7161–7165.

    PubMed  CAS  Google Scholar 

  7. Fromm, M. L., Taylor, P., and Walbot, V. (1986) Stable transformation of maize after gene transfer by electroporation. Nature 319, 791–793.

    PubMed  CAS  Google Scholar 

  8. Calvin, N. M. and Hanawalt, P. C. (1988) High-efficiency transformation of bacterial cells by electroporation. J. Bacteriol. 170, 2796–2801.

    PubMed  CAS  Google Scholar 

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

    PubMed  CAS  Google Scholar 

  10. Dower, W. J., Chassy, B. M., Trevors, J. T., and Blaschek, H. P. (1992) Protocols for the transformation of bacteria by electroporation, in Guide to Electroporation and Electrofusion (Chang, D. C., Chassy, B. M., Saunders, J. A., and Sowers, A. E., eds.), Academic, San Diego, pp 485–499.

    Google Scholar 

  11. Trevors, J. T., Chassy, B. M., Dower, W. J., and Blaschek, H. P. (1992) Electrotransformation of bacteria by plasmid DNA, in Guide to Electroporation and Electrofusion (Chang, D. C., Chassy, B. M., Saunders, J. A., and Sowers, A. E., eds.), Academic, San Diego, pp. 265–290.

    Google Scholar 

  12. Smith, M., Jessee, J., Landers, T., and Jordan, J. (1990) High efficiency bacterial electroporation: 1 × 1010 E. coli transformants/µg. Focus 12, 38–40.

    Google Scholar 

  13. Hanahan, D. (1983) Studies on transformation of Escherichia coli with plasmids. J. Mol. Biol. 166, 557–580.

    PubMed  CAS  Google Scholar 

  14. Huynh, T. V., Young, R. A., and Davis, R. W. (1985) Constructing and screening cDNA libraries in λgt10 and λgt11, in DNA Cloning: A Practical Approach (Glover, D. M., ed.) IRL, Oxford, pp. 49–78.

    Google Scholar 

  15. Hanahan, D. (1985) Techniques for transformation of E. coli, in DNA Cloning: A Practical Approach (Glover, D. M., ed.) IRL, Oxford, pp. 109–135.

    Google Scholar 

  16. 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 

  17. Rubenstein, J. L., Bruce, A. J., Ciaranello, R. D., Denney, D., Porteus, M. H., and Usdin, T. B. (1990) Subtractive hybridization system using single-stranded phagemids with directional inserts. Nucleic Acids Res. 18, 4833–4842.

    PubMed  CAS  Google Scholar 

  18. Li, W.-B., Gruber, C. E., Lin, J.-J., Lim, R., D’Alessio, J. M., and Jessee, J. A. (1994) The isolation of differentially expressed genes in fibroblast growth factor stimulated BC3H1 cells by subtractive hybridization. BioTechniques 16, 722–729.

    PubMed  CAS  Google Scholar 

  19. Chassy, B. M. and Flickinger, J. L. (1987) Transformation of Lactobacillus casei by electroporation. FEMS Microbiol. Lett. 44, 173–177.

    CAS  Google Scholar 

  20. Fiedler, S. and Wirth, R. (1988) Transformation of bacteria with plasmid DNA by electroporation. Anal. Biochem. 170, 38–44.

    PubMed  CAS  Google Scholar 

  21. Dunny, G. M., Lee, L. N., and LeBlanc, D. J. (1991) Improved electroporation and cloning vector system for gram-positive bacteria. Appl Environ Microbiol. 57, 1194–1201.

    PubMed  CAS  Google Scholar 

  22. Dower, W. J. (1990) Electroporation of bacteria: a general approach to genetic transformation, in Genetic Engineering—Principles and Methods, vol. 12 (Setlow, J. K., ed.) Plenum, New York, pp. 275–296.

    Google Scholar 

  23. Kotewicz, M. L., Sampson, C. M., D’Alessio, J. M., and Gerard, G. F. (1988) Isolation of cloned Moloney murine leukemia virus reverse transcriptase lacking ribonuclease H activity. Nucleic Acids Res 16, 265–277.

    PubMed  CAS  Google Scholar 

  24. Gerard, G. F., D’Alessio, J. M., and Kotewicz, M. L. (1989) cDNA synthesis by cloned Moloney murine leukemia virus reverse transcriptase lacking RNase H activity. Focus 11, 66–69.

    Google Scholar 

  25. D’Alessio, J. M., Gruber, C. E., Cain, C., and Noon, M. C. (1990) Construction of directional cDNA libraries using the SUPERSCRIPT Plasmid System. Focus 12, 47–50.

    Google Scholar 

  26. Willson, T. A. and Gough, N. M. (1988) High voltage E. coli electrotransformation with DNA following ligation. Nucleic Acids Res. 16, 11,820.

    PubMed  CAS  Google Scholar 

  27. Heery, D. M. and Dunican, L. K. (1989) Improved efficiency M13 cloning using electroporation. Nucleic Acids Res. 17, 8006.

    PubMed  CAS  Google Scholar 

  28. Jacobs, M., Wnendt, S., and Stahl, U. (1990) High-efficiency electrotransformation of Escherichia coli with DNA from ligation mixtures. Nucleic Acids Res. 18, 1653.

    PubMed  CAS  Google Scholar 

  29. Kobori, M. and Nojima, H. (1993) A simple treatment of DNA in a ligation mixture prior to electroporation improves transformation frequency. Nucleic Acids Res. 21, 2782.

    PubMed  CAS  Google Scholar 

  30. Zabarovsky, E. R. and Winberg, G. (1990) High efficiency electroporation of ligated DNA into bacteria. Nucleic Acids Res. 18, 5912.

    PubMed  CAS  Google Scholar 

  31. Taketo, A. (1988) DNA transfection of Escherichia coli by electroporation. Biochim. Biophys. Acta 949, 318–324.

    PubMed  CAS  Google Scholar 

  32. Leonardo, E. D. and Sedivy, J. M. (1990) A new vector for cloning large eukaryotic DNA segments in Escherichia coli. BioTechnology 8, 841–844.

    PubMed  CAS  Google Scholar 

  33. Lin, J.-J., Jessee, J., and Bloom, F. (1992) DH12S: a new electrocompetent E. coli strain for the production of highly purified single-stranded DNA using phagemid vectors. Focus 14, 98–101.

    Google Scholar 

  34. Siguret, V., Ribba, A.-S., Cherel, G., Meyer, D., and Pietu, G. (1994) Effect of plasmid size on transformation efficiency by electroporation of Escherichia coli DH5α. BioTechniques 16, 422–426.

    PubMed  CAS  Google Scholar 

  35. Hanahan, D., Jessee, J., and Bloom, F. (1991) Plasmid transformation of Escherichia coli and other bacteria, in Methods in Enzymology, vol. 204, Academic, San Diego, CA, pp. 63–113.

    Google Scholar 

  36. Gruber, C. E. (1992) High-efficiency cDNA cloning: a comparison of electroporation and in vitro packaging. BioTechniques 12, 804–808.

    PubMed  CAS  Google Scholar 

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Author information

Authors and Affiliations

  1. Research and Development, Life Technologies, Gaithersburg, MD

    Christian E. Gruber

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  1. Christian E. Gruber
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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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Gruber, C.E. (1995). Production of cDNA Libraries by Electroporation. In: Nickoloff, J.A. (eds) Electroporation Protocols for Microorganisms. Methods in Molecular Biology™, vol 47. Humana Press. https://doi.org/10.1385/0-89603-310-4:67

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  • DOI: https://doi.org/10.1385/0-89603-310-4:67

  • 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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