Advertisement

Analysis of Cytomegalovirus Gene Expression by Transfection

Protocol
  • 255 Downloads
Part of the Methods in Molecular Medicine™ book series (MIMM, volume 33)

Abstract

Human cytomegalovirus (HCMV), a human herpesvirus, is the leading viral cause of birth defects (1,2). In acquired immune deficiency syndrome (AIDS) patients, HCMV can cause severely debilitating colitis and retinitis; it is now an increasingly common cause of life-threatening pneumonitis in transplant patients (3, 4, 5). HCMV has a severely restricted host range in culture. It grows efficiently only in primary human diploid fibroblasts (HFF) and a few selected human cell lines (6, 7, 8, 9). Monocytes serve as a major reservoir of latent HCMV in humans and HCMV can be grown in vitro in primary monocyte-derived macrophages (M/M) (10, 11, 12, 13, 14).

Keywords

HeLa Cell Acquire Immune Deficiency Syndrome DEAE Dextran Luciferase Assay Reagent Restricted Host Range 
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.
This is a preview of subscription content, log in to check access.

References

  1. 1.
    Stagno, S., Pass, R. F., and Alford, C. A. (1981) Perinatal infections and maldevelopment. Birth Defects 17, 31–50.PubMedGoogle Scholar
  2. 2.
    Britt, W. J. and Alford, C. A. (1996) Cytomegalovirus, in Fields’ Virology, 3rd ed. (Fields, B. N., Knipe, D. M., and Howley, P. M., eds.), Lippincott-Raven Press, Philadelphia, pp. 2493–2523.Google Scholar
  3. 3.
    Ho, M. (1995) Cytomegalovirus, in Principles and Practice of Infectious Diseases (Mandell, G. L., Bennett, J. E., and Dolin, R., eds.), Churchill Livingstone, New York, Edinburgh, London, Melbourne, Tokyo, pp. 1351–1364.Google Scholar
  4. 4.
    Rubin, R. H. (1990) Impact of cytomegalovirus infection on organ transplant recipients. Rev. Infect. Dis. 12S7, 754–766.Google Scholar
  5. 5.
    Winston, D. J., Ho, W. G., and Champlin, R. E. (1990) Cytomegalovirus infections after allogeneic bone marrow transplantation. Rev. Infect. Dis. 12S7, S776–S792.Google Scholar
  6. 6.
    Gönczöl, E., Andrews, P. W., and Plotkin, S. A. (1984) Cytomegalovirus replicates in differentiated but not in undifferentiated human embryonal carcinoma cells. Science 224, 159–161.CrossRefPubMedGoogle Scholar
  7. 7.
    Smith, J. D. (1986) Human cytomegalovirus: demonstration of permissive epithelial cells and nonpermissive fibroblastic cells in a survey of human cell lines. J. Virol. 60, 583–588.PubMedGoogle Scholar
  8. 8.
    Compton, T. (1993) An immortalized human fibroblast cell line is permissive for human cytomegalovirus infection. J. Virol. 67, 3644–3648.PubMedGoogle Scholar
  9. 9.
    Sinzger, C. and Jahn, G. (1996) Human cytomegalovirus cell tropism and pathogenesis. Intervirology 39, 302–319.PubMedGoogle Scholar
  10. 10.
    Taylor-Wiedeman, J., Sissons, P., and Sinclair, J. (1994) Induction of endogenous human cytomegalovirus gene expression after differentiation of monocytes from healthy carriers. J. Virol. 68, 1597–1604.PubMedGoogle Scholar
  11. 11.
    Kondo, K., Xu, J., and Mocarski, E. S. (1996) Human cytomegalovirus latent gene expression in granulocyte-macrophage progenitors in culture and in seropositive individuals. Proc. Natl. Acad. Sci. USA 93, 11,137–11,142.CrossRefPubMedGoogle Scholar
  12. 12.
    Sinclair, J. and Sissons, P. (1996) Latent and persistent infections of monocytes and macrophages. Intervirology 39, 293–301.PubMedGoogle Scholar
  13. 13.
    Soderberg-Naucler, C., fish, K. N., and Nelson, J. A. (1997) Reactivation of latent human cytomegalovirus by allogenic stimulation of blood cells from healthy donors. Cell 91, 119–126.CrossRefPubMedGoogle Scholar
  14. 14.
    Lathey, J. L. and Spector, S. A. (1991) Unrestricted replication of human cytomegalovirus in hydrocortisone-treated macrophages. J. Virol. 65, 6371–6375.PubMedGoogle Scholar
  15. 15.
    Colberg-Poley, A. M., Santomenna, L. D., Harlow, P. P., Benfield, P. A., and Tenney, D. J. (1992) The HCMV US3 and UL36–38 immediate early proteins regulate gene expression. J. Virol. 66, 95–105.PubMedGoogle Scholar
  16. 16.
    Colberg-Poley, A. M., Huang, L., Soltero, V. F., Iskenderian, A., Schumacher, R.-F., and Anders, D. G. (1998) The acidic domain of pμL37x1 and gp μL37 plays a keyrole in transactivation of HCMV DNA replication gene promoters. Virology 246, 400–408.CrossRefPubMedGoogle Scholar
  17. 17.
    Zhang, H., Al-Barazi, H. O., and Colberg-Poley, A. M. (1996) The acidic domain of the human cytomegalovirus UL37 immediate early protein is dispensable for its transactivating activity and localization but is not for its synergism. Virology 223, 292–302.CrossRefPubMedGoogle Scholar
  18. 18.
    Aldovini, A. and Feinberg, M. B. (1990) Transfection of molecularly cloned HIV genomes, in Techniques in HIV Research (Aldovini, A. and Walker, B. D.), Stockton Press, New York, pp. 147–175.Google Scholar
  19. 19.
    Jones, T. R., Muzithras, V. P., and Gluzman, Y. (1991) Replacement mutagenesis of the human cytomegalovirus genome: US10 and US11 gene products are nonessential. J. Virol. 65, 5860–5872.PubMedGoogle Scholar
  20. 20.
    Kemble, G., Duke, G., Winter, R., and Spaete, R. (1996) Defined large-scale alteration of the human cytomegalovirus genome constructed by cotransfection of overlapping cosmids. J. Virol. 70, 2044–2048.PubMedGoogle Scholar
  21. 21.
    Baldick, C. J., Jr., Marchini, A., Patterson, C. E., and Shenk, T. (1997) Human cytomegalovirus tegument protein pp71 (ppUL82) enhances the infectivity of viral DNA and accelerates the infectious cycle. J. Virol. 71, 4400–4408.PubMedGoogle Scholar
  22. 22.
    Iskenderian, A. C., Huang, L., Reilly, A., Stenberg, R. M., and Anders, D. G. (1996) Four of eleven loci required for transient complementation of human cytomegalovirus DNA replication cooperate to activate expression of replication genes. J. Virol. 70, 383–392.PubMedGoogle Scholar
  23. 23.
    Eustice, D. C., Feldman, P. A., Colberg-Poley, A. M., Buckery, R. M., Neubauer, R. H. (1991) A sensitive method for detection of β-galactosidase in transfected mammalian cells. BioTechniques 11, 739–743.PubMedGoogle Scholar
  24. 24.
    Al-Barazi, H. O. and Colberg-Poley, A. M. (1996) The human cytomegalovirus UL37 immediate-early regulatory protein is an integral membrane N-glycoprotein which traffics through the endoplasmic reticulum and Golgi apparatus. J. Virol. 70, 7198–7208.PubMedGoogle Scholar
  25. 25.
    DeWet, J. R., Wood, K. V., DeLuca, M., Helinski, D. R., and Subramani, S. (1987) Firefly luciferase gene: structure and expression in mammalian cells. Mol. Cell. Biol. 7, 725–737.Google Scholar

Copyright information

© Humana Press Inc. 2000

Authors and Affiliations

  1. 1.Center for Immunology and Infectious Disease ResearchChildren’s National Medical CenterWashington, DC

Personalised recommendations

Cite protocol

Buy options