Manufacturing Viruses

  • Milton W. TaylorEmail author


The manufacturing of viruses in the test tube begins with the observations that viral capsid proteins undergo self-assembly when ionic and pH conditions are suitable. Components of Escherichia coli phage, such as T4 and λ, assemble spontaneously to form whole phage particles, and components of icosahedral viruses in the presence or absence of nucleic acid form spherical structures. Viral RNA was copied by the reverse transcriptase into cDNA and expressed on a plasmid with the formation of intact virus. By 2002, poliovirus was synthesized from full-length cDNA. With the progress in technology for the synthesis of long stretches of DNA and improvements in sequencing, it was a natural next step to construct a virus in a cell-free system using commercially available DNA. Poliovirus and Qβ bacteriophage were synthesized in vitro; similar techniques were used to sequence and reconstitute the 1918 flu pandemic virus, and to modify the host range of avian influenza virus. These latter experiments were criticized as being dangerous to humans and to the environment, and new regulations were introduced to control such experiments.


Influenza Virus H5N1 Virus Avian Influenza Virus Yersinia Pestis Francisella Tularensis 
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  1. 1.
    Cello, J., Paul, A. V., & Wimmer, E. (2002). Chemical synthesis of poliovirus cDNA: generation of infectious virus in the absence of natural template. Science, 297(5583), 1016–1018.PubMedCrossRefGoogle Scholar
  2. 2.
    Edgar, R. S., & Lielausis, I. (1968). Some steps in the assembly of bacteriophage T4. Journal of Molecular Biology, 32(2), 263–276.PubMedCrossRefGoogle Scholar
  3. 3.
    Weigle, J. (1966). Assembly of phage lambda in vitro. Proceedings of the National Academy of Sciences of the United States of America, 55(6), 1462–1466.PubMedCentralPubMedGoogle Scholar
  4. 4.
    Putnak, J. R., & Phillips, B. A. (1981). Picornaviral structure and assembly. Microbiological Reviews, 45(2), 287–315.PubMedCentralPubMedGoogle Scholar
  5. 5.
    Bancroft, J. B., & Hiebert, E. (1967). Formation of an infectious nucleoprotein from protein and nucleic acid isolated from a small spherical virus. Virology, 32(2), 354–356.PubMedCrossRefGoogle Scholar
  6. 6.
    Bancroft, J. B., Hills, G. J., & Markham, R. (1967). A study of the self-assembly process in a small spherical virus. Formation of organized structures from protein subunits in vitro. Virology, 31(2), 354–379.PubMedCrossRefGoogle Scholar
  7. 7.
    Fraenkel-Conrat, H., & Singer, B. (1957). Virus reconstitution. II. Combination of protein and nucleic acid from different strains. Biochimica et Biophysica Acta, 24(3), 540–548.PubMedCrossRefGoogle Scholar
  8. 8.
    Taniguchi, T., Palmieri, M., & Weissmann, C. (1978). QB DNA-containing hybrid plasmids giving rise to QB phage formation in the bacterial host. Nature, 274(5668), 223–228.PubMedCrossRefGoogle Scholar
  9. 9.
    Racaniello, V. R., & Baltimore, D. (1981). Cloned poliovirus complementary DNA is infectious in mammalian cells. Science, 214(4523), 916–919.PubMedCrossRefGoogle Scholar
  10. 10.
    Racaniello, V. R., & Baltimore, D. (1981). Molecular cloning of poliovirus cDNA and determination of the complete nucleotide sequence of the viral genome. Proc Natl Acad Sci USA, 78(8), 4887–4891.PubMedCentralPubMedCrossRefGoogle Scholar
  11. 11.
    Wimmer, E., Mueller, S., Tumpey, T. M., & Taubenberger, J. K. (2009). Synthetic viruses: a new opportunity to understand and prevent viral disease. Nature Biotechnology, 27(12), 1163–1172.PubMedCentralPubMedCrossRefGoogle Scholar
  12. 12.
    Agarwal, K. L., Buchi, H., Caruthers, M. H., Gupta, N., Khorana, H. G., Kleppe, K., et al. (1970). Total synthesis of the gene for an alanine transfer ribonucleic acid from yeast. Nature, 227(5253), 27–34.PubMedCrossRefGoogle Scholar
  13. 13.
    Caruthers, M. H., Kleppe, R., Kleppe, K., & Khorana, H. G. (1976). Total synthesis of the structural gene for the precursor of a tyrosine suppressor transfer RNA from Escherichia coli. 10. Enzymatic joining of chemically synthesized segments to form the DNA duplex corresponding to the nucleotide sequence 86-126. Journal of Biological Chemistry, 251(3), 658–666.PubMedGoogle Scholar
  14. 14.
    Khorana, H. G. (1979). Total synthesis of a gene. Science, 203(4381), 614–625.PubMedCrossRefGoogle Scholar
  15. 15.
    De Jesus, N., Franco, D., Paul, A., Wimmer, E., & Cello, J. (2005). Mutation of a single conserved nucleotide between the cloverleaf and internal ribosome entry site attenuates poliovirus neurovirulence. Journal of Virology, 79(22), 14235–14243.PubMedCentralPubMedCrossRefGoogle Scholar
  16. 16.
    Smith, H. O., Hutchison, C. A, 3rd, Pfannkoch, C., & Venter, J. C. (2003). Generating a synthetic genome by whole genome assembly: phiX174 bacteriophage from synthetic oligonucleotides. Proceedings of the National Academy of Sciences of the United States of America, 100(26), 15440–15445.PubMedCentralPubMedGoogle Scholar
  17. 17.
    Herfst, S., Schrauwen, E. J., Linster, M., Chutinimitkul, S., de Wit, E., Munster, V. J., et al. (2012). Airborne transmission of influenza A/H5N1 virus between ferrets. Science, 336(6088), 1534–1541.PubMedCrossRefGoogle Scholar
  18. 18.
    Chutinimitkul, S., van Riel, D., Munster, V. J., van den Brand, J. M., Rimmelzwaan, G. F., Kuiken, T., et al. (2010). In vitro assessment of attachment pattern and replication efficiency of H5N1 influenza A viruses with altered receptor specificity. Journal of Virology, 84(13), 6825–6833.PubMedCentralPubMedCrossRefGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2014

Authors and Affiliations

  1. 1.Indiana UniversityBloomingtonUSA

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