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DNA Affinity Purification of Epstein-Barr Virus OriP-Binding Proteins

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DNA Viruses

Part of the book series: Methods in Molecular Biology ((MIMB,volume 292))

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Abstract

DNA affinity purification has been used to identify cellular and viral proteins associated with the Epstein-Barr virus origin of plasmid DNA replication. This approach allows for a one- or two-step purification scheme of high-affinity DNA binding proteins from crude nuclear extracts. Additionally, this approach may be useful for isolation of proteins that are found in the insoluble fractions of the nuclear matrix or scaffold.

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References

  1. Kadonaga, J. T. and Tjian, R. (1986) Affinity purification of sequence-specific DNA binding proteins. Proc. Natl. Acad. Sci. USA 83, 5889–5893.

    Article  PubMed  CAS  Google Scholar 

  2. Rosenfeld, P. J. and Kelly, T. J. (1986) Purification of nuclear factor I by DNA recognition site affinity chromatography. J. Biol. Chem. 261, 1398–1408.

    PubMed  CAS  Google Scholar 

  3. Gadgil, H., Oak, S. A., and Jarrett, H. W. (2001) Affinity purification of DNA-binding proteins. J. Biochem. Biophys. Methods 49, 607–624.

    Article  PubMed  CAS  Google Scholar 

  4. Kadonaga, J. T. (1991) Purification of sequence-specific binding proteins by DNA affinity chromatography. Methods Enzymol. 208, 10–23.

    Article  PubMed  CAS  Google Scholar 

  5. Ranish, J. A., Yudkovsky, N., and Hahn, S. (1999) Intermediates in formation and activity of the RNA polymerase II preinitiation complex: holoenzyme recruitment and a postrecruitment role for the TATA box and TFIIB. Genes Dev. 13, 49–63.

    Article  PubMed  CAS  Google Scholar 

  6. Kumar, R., Reynolds, D. M., Shevchenko, A., Shevchenko, A., Gladstone, S. D., Dalton, S. (2000) Forkhead transcription factors, Fkh1p and Fkh2p, collaborate with Mcm1p to control transcription required for M-phase. Curr. Biol. 10, 896–906.

    Article  PubMed  CAS  Google Scholar 

  7. Sugden, B. and Leight, E. R. (2001) Molecular mechanisms of maintenance and disruption of virus latency, in Epstein-Barr Virus and Human Cancer (Takada, K., ed.), Springer, Heidelberg, pp. 3–11.

    Google Scholar 

  8. Leight, E. R. and Sugden, B. (2000) EBNA-1: a protein pivotal to latent infection by Epstein-Barr virus. Rev. Med. Virol. 10, 83–100.

    Article  PubMed  CAS  Google Scholar 

  9. Yates, J. L. Warren, N., Reisman, D., Sugden, B., et al. (1984) A cis-acting element from Epstein-Barr viral genome that permits stable replication of recombinant plasmids in latently infected cells. Proc. Natl. Acad. Sci. USA 81, 3806–3810.

    Article  PubMed  CAS  Google Scholar 

  10. Rawlins, D. R. Millman, G., Hayward, S. D., Howard, G. S. (1985) Sequence-specific DNA binding of the Epstein-Barr virus nuclear antigen (EBNA-1) to clustered sites in the plasmid maintenance region. Cell 42, 859–868.

    Article  PubMed  CAS  Google Scholar 

  11. Dignam, J. D., Lebovitz, R. M., and Roeder, R. G. (1983) Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 11, 1475–1489.

    Article  PubMed  CAS  Google Scholar 

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© 2005 Humana Press Inc.

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Atanasiu, C., Lezina, L., Lieberman, P.M. (2005). DNA Affinity Purification of Epstein-Barr Virus OriP-Binding Proteins. In: Lieberman, P.M. (eds) DNA Viruses. Methods in Molecular Biology, vol 292. Humana Press. https://doi.org/10.1385/1-59259-848-X:267

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  • DOI: https://doi.org/10.1385/1-59259-848-X:267

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-353-4

  • Online ISBN: 978-1-59259-848-9

  • eBook Packages: Springer Protocols

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