Virus Genes

, Volume 12, Issue 2, pp 193–197 | Cite as

Enhancement of HSV-DNA infectivity, in Vero and RS cells, by a modified calcium-phosphate transfection technique

  • Mauro Tognon
  • Elisa Margherita Cattozzo
  • Stefano Bianchi
  • Maria Grazia Romanelli
Article

Abstract

Infectivity of herpes simplex virus (HSV) DNA was assessed by employing the calciumphosphate transfection technique described by Chen and Okayama, originally applied to increase the efficiency of plasmid transfection by N,N-bis (2-hydroxyethyl)-2-aminoethane sulfonic acid (BES). The experimental conditions and efficiency of this transfection procedure were evaluated comparing the viral progeny titers obtained by the Chen and Okayama transfection method using DNA from wild-type strains of HSV-1 and HSV-2, as well as from mutant strains, with the viral progeny obtained by the most widely used transfection technique introduced by Graham and van der Eb. Furthermore, recombinant virus production was evaluated in marker transfer and marker rescue experiments, comparing both transfection techniques, using DNA fragments cotransfected with whole viral DNA into African green monkey (Vero) or rabbit skin (RS) cells. The viral production obtained from HSV-DNA transfected cells was enhanced approximately 1000-fold when the Chen and Okayama procedure was applied.

Key words

herpes simplex virus DNA calcium-phosphate transfection infectivity assay eukaryotic cell recombinant virus 

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References

  1. 1.
    GrahamF.L., VeldhuisenG., and WilkieN.M., Nature New Biol245, 265–266, 1973.Google Scholar
  2. 2.
    StowN.D. and WilkieN.M., J Gen Virol33, 447–458, 1976.Google Scholar
  3. 3.
    TognonM., FurlongD., ConleyA.J., and RoizmanB., J Virol40, 870–880, 1981.Google Scholar
  4. 4.
    GrahamF.L. and van derEbA.J., Virology52, 456–467, 1973.Google Scholar
  5. 5.
    VaheriA. and PaganoJ.S., Virology27, 435–442, 1965.Google Scholar
  6. 6.
    ParkerB.A. and StrakG.A., J Virol31, 360–369, 1979.Google Scholar
  7. 7.
    NeumannE., Schaefer-RidderM., WangY., and HofschneiderP.H., EMBO J1, 641–845, 1982.Google Scholar
  8. 8.
    CameronI.R., WilkieN.M., and MacnabJ.C.M., J Virol Methods6, 183–191, 1983.Google Scholar
  9. 9.
    BondV.C. and WoldB., Mol Cell Biol7, 2286–2293, 1987.Google Scholar
  10. 10.
    BrashD.E., ReddelR.R., QuanrudM., YangK., FarrellM.P., and HarrisC.C., Mol Cell Biol7, 2031–2034, 1987.Google Scholar
  11. 11.
    JackersP., DefechereuxP., BaudouxL., LambertC., MassaerM., Merville-LouisM.P., RentierB., and PietteJ., J Virol66, 3899–3903, 1992.Google Scholar
  12. 12.
    ChenC. and OkayamaH., Mol Cell Biol7, 2745–2752, 1987.Google Scholar
  13. 13.
    TognonM., GuandaliniR., RomanelliM.G., ManservigiR., and TrevisaniB., Virus Res18, 135–150, 1991.Google Scholar
  14. 14.
    RomanelliM.G., CattozzoE.M., FaggioliL., and TognonM., J Gen Virol72, 1991–1995, 1991.Google Scholar
  15. 15.
    PostL.E., MackemS., and RoizmanB., Cell24, 555–565, 1981.Google Scholar
  16. 16.
    CassaiE., MenariniL., and TerniM., Arch Vet Ital23, 183–186, 1972.Google Scholar
  17. 17.
    CourtneyR.J. and PowellK.L., in de-TheG., EpsteinM.A., and Zur HausenH. (eds).Proceedings of the Symposium on Herpesviruses and Oncogenesis. International Agency for Research Against Cancer, Lyon, France, 1975, pp 63–73.Google Scholar
  18. 18.
    EjercitoP.M., KieffE.D., and RoizmanB., J Gen Virol2, 357–364, 1968.Google Scholar
  19. 19.
    WalboomersJ.M. and ter SchaggetJ., Virology74, 256–258, 1976.Google Scholar

Copyright information

© Kluwer Academic Publishers 1996

Authors and Affiliations

  • Mauro Tognon
    • 1
  • Elisa Margherita Cattozzo
    • 1
  • Stefano Bianchi
    • 1
  • Maria Grazia Romanelli
    • 2
  1. 1.Institute of Histology and General Embryology, School of MedicineUniversity of FerraraFerraraItaly
  2. 2.Institute of Biology and Genetics, School of MedicineUniversity of VeronaVeronaItaly

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