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Experimental Strategies in Efficient Transfection of Mammalian Cells

Calcium Phosphate and DEAE-Dextran
  • Gregory S. Pari
  • Wayne A. Keown
Part of the Methods in Molecular Biology book series (MIMB, volume 62)

Abstract

Transfection of foreign DNA into mammalian cells is one of the most common procedures performed by investigators next to cell culture itself. Whether the goal is to express mammalian or viral genes transiently or by the generation of stable cells lines, researchers have a variety of transfection methods to choose from Calcium phosphate coprecipitation and DEAE-dextran-mediated transfection are the oldest, and with many investigators, the most trusted of these methods. Paradoxically, the exact mechanism of DNA uptake for each of these methods remains unknown.

Keywords

Calcium Phosphate Express Cell Line Calcium Phosphate Method Effector Plasmid Calcium Phosphate Transfection 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Fujita, T., Shubiya, H., Ohashi, T., Yamanishi, K., and Taniguchi, T. (1986) Regulation of human interleukin-2 gene. Functional DNA sequences in the 5′ flanking region for the gene expression in activated T lymphocytes. Cell 46, 401–407.PubMedCrossRefGoogle Scholar
  2. 2.
    Lopata, M. A., Cleveland, D. W., and Sollner-Webb, B. (1984) High-level expression of a chloramphenicol acetyltransferase gene by DEAE-dextran-mediated DNA transfection coupled with a dimethysulfoxide or glycerol shock treatment. Nucleic Acids Res. 12, 5707–5711.PubMedCrossRefGoogle Scholar
  3. 3.
    Reeves, R., Gorman, C., and Howard, B. (1985) Minichromosomes assembly of nonintegrated plasmid DNA transfected into mammalian cells. Nucleic Acids Res. 13, 3599–3605.PubMedCrossRefGoogle Scholar
  4. 4.
    Selden, R. F., Burke-Howie, K., Rowe, M. E., Goodman, H. M., and Moore, D. D. (1986) Human growth hormone as a reporter gene in regulation studies employing transient gene expression. Mol. Cell. Biol. 6, 3173–3179.PubMedGoogle Scholar
  5. 5.
    Sussman, D. J. and Milman, G. (1984) Short-term, high-efficiency expression of transfected DNA. Mol. Cell. Biol. 4, 1641–1646.PubMedGoogle Scholar
  6. 6.
    Pari, G. S. and Anders, D. G. (1993) Eleven loci encoding trans-acting factors are required for transient complementation of human cytomegalovirus oriLyt-dependent DNA replication. J. Virol. 67, 6979–6988.PubMedGoogle Scholar
  7. 7.
    Pari, G. S., Kacica, M. A., and Anders, D. A. (1993) Open reading frames UL44, IRS1/TRS1, and UL36-38 are required for transient complementation of human cytomegalovirus oriLyt-dependent DNA synthesis. J. Virol. 67, 2575–2582.PubMedGoogle Scholar
  8. 8.
    Graham, F. L. and Van der Eb, A. J. (1973) A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52, 456–460.PubMedCrossRefGoogle Scholar
  9. 9.
    Wigler, M., Pellicer, A., Silverstein, S., and Axel, R. (1978) Biochemical transfer of single-copy eucaryotic genes using total cellular DNA as donor. Cell 14, 725–731.PubMedCrossRefGoogle Scholar
  10. 10.
    Chen, C. and Okayama, H. (1987) High-efficiency transformation of mammalian cells by plasmid DNA. Mol. Cell. Biol. 7, 2745–2752.PubMedGoogle Scholar

Copyright information

© Humana Press Inc. 1997

Authors and Affiliations

  • Gregory S. Pari
    • 1
  • Wayne A. Keown
    • 1
  1. 1.HybridonWorcester

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