Growth and Characterization of Poliovirus Antigen Chimeras

  • David J. Evans
  • Philip D. Minor
Part of the Methods in Molecular Biology book series (MIMB, volume 8)


 Chapter 22 outlines the construction of engineered, full-length poliovirus cDNAs in which the region encoding a well-characterized antigenic site has been replaced by sequences of choice. This chapter briefly describes the methods used to generate, maintain, and characterize infectious chimeric viruses. These techniques include several developed during the course of poliovirus study that have recently been published (1). This overview describes modifications to these techniques where relevant, but concentrates on methods not detailed by Minor (1). The production and analysis of poliovirus antigen chimeras can be readily subdivided into three discrete areas.


Antigenic Site Newborn Calf Serum Chimeric Virus Cesium Chloride Routine Growth 
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.


  1. 1.
    Minor, P. D. (1985) Growth, assay and purification of picornaviruses, in Virology: A Practical Approach (IRL Press, Oxford, UK), pp. 25–41.Google Scholar
  2. 2.
    van der Werf, S., Bradley, J., Wimmer, E., Studier, F. W., and Dunn, J. J. (1986) Synthesis of infectious poliovirus RNA purified T7 RNA polymerase. Proc. Natl. Acad. Sci. USA 83, 2330–2334.PubMedCrossRefGoogle Scholar
  3. 3.
    Birnboim, H. C. and Doly, J. (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 7, 1513–1523.PubMedCrossRefGoogle Scholar
  4. 4.
    Saiki, R. K, Gelfand, D. H., Stoffel, S., Scharf, S.J., Higuchi, R., Horn, G. T., Mullis, K B., and Erlich, H. A. (1988) Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239, 487–491.PubMedCrossRefGoogle Scholar
  5. 5.
    Rico-Hesse, R., Pallansch, M. A., Nottay, B. K, and Kew, O. M. (1987) Geographic distribution of wild poliovirus type 1 genotypes. Virology 160, 311–322.PubMedCrossRefGoogle Scholar
  6. 6.
    Sanger, F., Nicklen, S., and Coulsen, A. R. (1977) DNA sequencing with chain-terminating inhibitors. Proc. Natl Acad. Sci. USA 74, 5463–5467.PubMedCrossRefGoogle Scholar
  7. 7.
    Nomoto, A., Omata, T., Toyada, H., Kuge, S., Hone, H, Katoaka, Y., Genba, Y., Nakano, Y, and Imura, N. (1982) Complete nucleotide sequence of the attenuated poliovirus Sabin 1 strain genome. Proc. Natl. Acad. Sci. USA 79, 5793–5797.PubMedCrossRefGoogle Scholar
  8. 8.
    Maniatis, T., Fritsch, E. F., and Sambrook, J. (1982) Molecular Cloning: A Laboratory Manual Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.Google Scholar
  9. 9.
    Winship, P. (1989) An improved method for directly sequencing PCR amplified material using dimethyl sulphoxide. Nucleic Acids Res. 17, 1266.PubMedCrossRefGoogle Scholar
  10. 10.
    Ferguson, M., Minor, P. D., Magrath, D.I., Qui, Y-H., Spitz, M., and Schild, G. C. (1984) Neutralization epitopes on poliovirus type 3 particles: An analysis using monoclonal antibodies. J. Gen. Virol. 65, 197–201.PubMedCrossRefGoogle Scholar
  11. 11.
    Burke, K L., Evans, D. J, Jenkins, O., D’Souza, E., and Almond, J. W. (1989) A cassette vector for the construction of antigen chimaeras of poliovirus. J. Gen. Virol. 70, 2475–2479.PubMedCrossRefGoogle Scholar
  12. 12.
    Evans, D. J., McKeating, J., Meredith, J., Burke, K. L., Katrak, K, John, A., Ferguson, M., Minor, P. D., Weiss, R. A., and Almond, J. W. (1989) An engineered poliovirus elicits broadly reactive HIV-1 neutralizing antibodies. Nature 339, 385–388.PubMedCrossRefGoogle Scholar
  13. 13.
    Jenkins, O., Cason, J., Burke, K. L., Lunny, D., Gillen, A., Patel, D., McCance, D., and Almond, J. W. (1989) An antigen chimaera of poliovirus induces antibodies against Human Papillomavirus type 16. J. Vtrol. 64, 1201–1206.Google Scholar
  14. 14.
    Kwok, S. and Higuchi, R. (1989) Avoiding false positives with PCR. Nature 339, 237,238.PubMedCrossRefGoogle Scholar

Copyright information

© The Humana Press Inc., Clifton, NJ 1991

Authors and Affiliations

  • David J. Evans
    • 1
  • Philip D. Minor
    • 2
  1. 1.Department of MicrobiologyUniversity of ReadingReadingUK
  2. 2.National Institute for Biological Standards and ControlHertfordshireUK

Personalised recommendations