Skip to main content

Effect of Cis-Located Human Satellite DNA on Electroporation Efficiency

  • Protocol
Animal Cell Electroporation and Electrofusion Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 48))

  • 1157 Accesses

Abstract

The introduction of exogenous DNA into mammalian cells can ultimately result in the integration of the transfected DNA into the host genome (1,2). Any genes containing appropriate expression signals will, in most cases, be expressed both prior to and after integration. However, stable expression of the transfected genes is not always observed (3,4). One reason this is believed to occur is because the site of integration of the foreign DNA is virtually random with respect to both the exogenous DNA and the particular human chromosome (5). Therefore, it can be expected that not only the expression, but also the stability and copy number of the transfected gene will depend on the integration sites and chromosomal flanking sequences.

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

Access this chapter

eBook
USD 39.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 54.99
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

References

  1. Heartlein, M. W., Knoll, J. H. M., and Latt, S. A. (1988) Chromosome instability associated with human alphoid DNA transfected into the Chinese hamster genome. Mol. Cell. Biol. 8, 3611–3618.

    PubMed  CAS  Google Scholar 

  2. Talarico, D., Peverali, A. F., Ginelli, E., Meneveri, R., Mondello, C., and Della Valle, G. (1988) Satellite DNA induces unstable expression of the adjacent herpes simplex virus tk gene cotransfected in mouse cells. Mol. Cell Biol. 8, 1336–1344.

    PubMed  CAS  Google Scholar 

  3. Butner, K., and Lo, C. W. (1986) Modulation of tk expression in mouse pericentromeric heterochromatin. Mol. Cell. Biol. 6, 4440–4449.

    PubMed  CAS  Google Scholar 

  4. Butner, K. A., and Lo, C. W. (1986) High frequency DNA rearrangements associated with mouse centromeric satellite DNA. J. Mol. Biol. 187, 547–556.

    Article  PubMed  CAS  Google Scholar 

  5. Murnane, J. P., Yezzi, M. J., and Young, B. R. (1990) Recombination events during integration of transfected DNA into normal human cells. Nucleic Acids Res. 18, 2733–2738.

    Article  PubMed  CAS  Google Scholar 

  6. Grigliatti, T. (1991) Position-effect variegation—an assay for nonhistone chromosomal proteins and chromatin assembly and modifying factors. Methods in Cell Biol. 35, 587–627.

    Article  CAS  Google Scholar 

  7. Beridze, T. (1986) Satellite DNA. Springer-Verlag, Berlin, Germany.

    Google Scholar 

  8. Wallenburg, J. C., Nepveu, A., and Chartrand, P. (1987) Integration of a vector containing rodent repetitive elements in the rat genome. Nucleic Acids Res. 15, 7849–7863.

    Article  PubMed  CAS  Google Scholar 

  9. Fouquet, C. and DuBow, M. S. (1992) Effect of cis-located human satellite DNA on the electroporation efficiency of neo and HSV-1 tk containing plasmids. Mut. Res. 284, 321–328.

    CAS  Google Scholar 

  10. Mulligan, R. C., and Berg, P. (1980) Expression of a bacterial gene in mammalian cells. Science 209, 1422–1427.

    Article  PubMed  CAS  Google Scholar 

  11. Potter, H., Selden, R. F., and Kingston, R. E. (1987) Transfection by electroporation, in Current Protocols in Molecular Biology (Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A., and Struhl, K., eds.), Greene Publishing Associates and Wiley-Interscience, Brooklyn, NY, pp. 9.3.1–9.5.6.

    Google Scholar 

  12. Goring, D. R., and DuBow, M. S. (1985) A cytotoxic effect associated with 9-(1,3-dihydroxy-2-propoxymethyl) guanine is observed during the selection for drug resistant human cells containing a single Herpesvirus thymidine kinase gene. Biochem. Biophys. Res. Commun. 133, 195–201.

    Article  PubMed  CAS  Google Scholar 

  13. Sol, K., Lapointe, M., Macleod, M., Nadeau, C., and DuBow, M. S. (1986) A cloned fragment of Hela DNA containing consensus sequences of satellite II and III DNA hybridizes with the Drosophila P-element and with the 1.8 kb family of human KpnI fragments. Biochim. Biophys. Acta. 868, 128–135.

    PubMed  CAS  Google Scholar 

  14. Rhim, J. S., Cho, H. Y., and Huebner, R. J. (1975) Non-producer human cells induced by murine sarcoma virus. Int. J. Cancer 15, 23–29.

    Article  PubMed  CAS  Google Scholar 

  15. Horita, A., and Weber, L. J. (1964) Skin penetrating property of drugs dissolved in dimethylsulfoxide (DMSO) and other vehicles. Life Sci. 3, 1389–1395.

    Article  PubMed  CAS  Google Scholar 

  16. Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Plasmid vectors, in Molecular Cloning, a Laboratory Manual, vol. 1, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp. 1.42–1.46.

    Google Scholar 

  17. Tolias, P. P., and DuBow, M. S. (1987) The amino terminus of the bacteriophage D108 transposase protein contains a two-component, sequence-specific, DNA-binding domain. Virology 157, 117–126.

    Article  PubMed  CAS  Google Scholar 

  18. Stubblefield, E. (1968) Synchronization methods for mammalian cell cultures, in Methods in Cell Physiology (Prescott, D. M., ed.), Academic, New York, pp. 25–43.

    Google Scholar 

  19. Chu, G., Hayakawa, H., and Berg P. (1987) Electroporation for the efficient transfection of mammalian cells with DNA. Nucleic Acids Res. 15, 1311–1326.

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  21. Hartman, S. C. and Mulligan, R. C. (1988) Two dominant-acting selectable markers for gene transfer studies in mammalian cells. Proc. Natl. Acad. Sci. USA 85, 8047–8051.

    Article  PubMed  CAS  Google Scholar 

  22. Artlet, P., Grannemann, R., Friel, J., Bartsch, J., and Hauser, H. (1991) The prokaryotic neomycin-resistance-encoding gene acts as a transcriptional silencer in eukaryotic cells. Gene 99, 249–254.

    Article  Google Scholar 

  23. Brisebois, J. J. and DuBow, M. S. (1993) Selection for spontaneous null mutations in a chromosomally integrated HSV-1 thymidine kinase gene yields deletions and a mutation caused by intragenic illegitimate recombination. Mut. Res. 287, 191–205.

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1995 Humana Press Inc.

About this protocol

Cite this protocol

Saint-Dic, D., DuBow, M.S. (1995). Effect of Cis-Located Human Satellite DNA on Electroporation Efficiency. In: Nickoloff, J.A. (eds) Animal Cell Electroporation and Electrofusion Protocols. Methods in Molecular Biology, vol 48. Humana Press. https://doi.org/10.1385/0-89603-304-X:199

Download citation

  • DOI: https://doi.org/10.1385/0-89603-304-X:199

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-304-7

  • Online ISBN: 978-1-59259-535-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics