1887

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Volume 85, Issue 11

Development of a reverse-genetics system for demonstrates that the small hydrophobic (SH) and attachment (G) genes are not essential for virus viability Free

Abstract

(APV) is a member of the genus of the subfamily . This study describes the development of a reverse-genetics system for APV. A minigenome system was used to optimize the expression of the nucleoprotein, phosphoprotein, M2 and large polymerase proteins when transfected into Vero cells under the control of the bacteriophage T7 promoter. Subsequently, cDNA was transcribed from the virion RNA to make a full-length antigenome, which was also cloned under the control of the T7 promoter. Transfection of the full-length genome plasmid, together with the plasmids expressing the functional proteins in the transcription and replication complex, generated APV in the transfected cells. The recombinant virus was passaged and was identified by cytopathic effect (CPE) that was typical of APV, the presence of a unique restriction-endonuclease site in the cDNA copy of the genome and immunofluorescence staining with anti-APV antibodies. Replacement of the full-length wild-type antigenome with one lacking the small hydrophobic (SH) protein and the attachment (G) genes generated a virus that grew more slowly and produced atypical CPE with syncytia much larger than those seen with wild-type virus.

  • Received:
  • Accepted:
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2004-11-01
2024-05-02
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References

  1. Ahmadian G., Randhawa J. S., Easton A. J. 2000; Expression of the ORF-2 protein of the human respiratory syncytial virus M2 gene is initiated by a ribosomal termination-dependent reinitiation mechanism. EMBO J 19:2681–2689 [CrossRef]
     [Google Scholar]
  2. Baxter-Jones C., Wilding G. P., Grant M. 1986; Immunofluorescence as a potential diagnostic method for turkey rhinotracheitis. Vet Rec 119:600–601
     [Google Scholar]
  3. Britton P., Green P., Kottier S., Mawditt K. L., Penzes Z., Cavanagh D., Skinner M. A. 1996; Expression of bacteriophage T7 RNA polymerase in avian and mammalian cells by a recombinant fowlpox virus. J Gen Virol 77:963–967 [CrossRef]
     [Google Scholar]
  4. Bukreyev A., Whitehead S. S., Murphy B. R., Collins P. L. 1997; Recombinant respiratory syncytial virus from which the entire SH gene has been deleted grows efficiently in cell culture and exhibits site-specific attenuation in the respiratory tract of the mouse. J Virol 71:8973–8982
     [Google Scholar]
  5. Cavanagh D., Barrett T. 1988; Pneumovirus-like characteristics of the mRNA and proteins of turkey rhinotracheitis virus. Virus Res 11:241–256 [CrossRef]
     [Google Scholar]
  6. Collins M. S., Gough R. E. 1988; Characterization of a virus associated with turkey rhinotracheitis. J Gen Virol 69:909–916 [CrossRef]
     [Google Scholar]
  7. Collins P. L., Hill M. G., Camargo E., Grosfeld H., Chanock R. M., Murphy B. R. 1995; Production of infectious human respiratory syncytial virus from cloned cDNA confirms an essential role for the transcription elongation factor from the 5′ proximal open reading frame of the M2 mRNA in gene expression and provides a capability for vaccine development. Proc Natl Acad Sci U S A 92:11563–11567 [CrossRef]
     [Google Scholar]
  8. Cook J. K. A. 2000; Avian rhinotracheitis. Rev Sci Tech 19:602–613
     [Google Scholar]
  9. Jin H., Cheng X., Traina-Dorge V. L., Park H. J., Zhou H., Soike K., Kemble G. 2003; Evaluation of recombinant respiratory syncytial virus gene deletion mutants in African green monkeys for their potential as live attenuated vaccine candidates. Vaccine 21:3647–3652 [CrossRef]
     [Google Scholar]
  10. Jones R. C. 1996; Avian pneumovirus infection: questions still unanswered. Avian Pathol 25:639–648 [CrossRef]
     [Google Scholar]
  11. Jones R. C., Baxter-Jones C., Wilding G. P., Kelly D. F. 1986; Demonstration of a candidate virus for turkey rhinotracheitis in experimentally inoculated turkeys. Vet Rec 119:599–600
     [Google Scholar]
  12. Kapczynski D. R., Sellers H. S. 2003; Immunization of turkeys with a DNA vaccine expressing either the F or N gene of avian metapneumovirus. Avian Dis 47:1376–1383 [CrossRef]
     [Google Scholar]
  13. Karron R. A., Buonagurio D. A., Georgiu A. F. 8 other authors 1997; Respiratory syncytial virus (RSV) SH and G proteins are not essential for viral replication in vitro : clinical evaluation and molecular characterization of a cold-passaged, attenuated RSV subgroup B mutant. Proc Natl Acad Sci U S A 94:13961–13966 [CrossRef]
     [Google Scholar]
  14. Levine S., Klaiber-Franco R., Paradiso P. R. 1987; Demonstration that glycoprotein G is the attachment protein of respiratory syncytial virus. J Gen Virol 68:2521–2524 [CrossRef]
     [Google Scholar]
  15. Ling R., Pringle C. R. 1988; Turkey rhinotracheitis virus: in vivo and in vitro polypeptide synthesis. J Gen Virol 69:917–923 [CrossRef]
     [Google Scholar]
  16. Ling R., Easton A. J., Pringle C. R. 1992; Sequence analysis of the 22K, SH and G genes of turkey rhinotracheitis virus and their intergenic regions reveals a gene order different from that of other pneumoviruses. J Gen Virol 73:1709–1715 [CrossRef]
     [Google Scholar]
  17. Marriott A. C., Easton A. J. 1999; Reverse genetics of the Paramyxoviridae . Adv Virus Res 53:321–340
     [Google Scholar]
  18. Naylor C. J., Jones R. C. 1994; Demonstration of a virulent subpopulation in a prototype live attenuated turkey rhinotracheitis vaccine. Vaccine 12:1225–1230 [CrossRef]
     [Google Scholar]
  19. Naylor C. J., Jones R. C., Cavanagh D. 2003; Trends in live avian pneumovirus vaccination in the UK. Some developments in live TRT vaccination in the UK. In Proceedings of the XIII Congress of the World Veterinary Poultry Association p– 67 July 19–23 2003 Denver, CO, USA:
     [Google Scholar]
  20. Peeters B. P. H., de Leeuw O. S., Koch G., Gielkens A. L. J. 1999; Rescue of Newcastle disease virus from cloned cDNA: evidence that cleavability of the fusion protein is a major determinant for virulence. J Virol 73:5001–5009
     [Google Scholar]
  21. Randhawa J. S., Marriott A. C., Pringle C. R., Easton A. J. 1997; Rescue of synthetic minireplicons establishes the absence of the NS1 and NS2 genes from avian pneumovirus. J Virol 71:9849–9854
     [Google Scholar]
  22. Schmidt U., Beyer J., Polster U., Gershwin L. J., Buchholz U. J. 2002; Mucosal immunization with live recombinant bovine respiratory syncytial virus (BRSV) and recombinant BRSV lacking the envelope glycoprotein G protects against challenge with wild-type BRSV. J Virol 76:12355–12359 [CrossRef]
     [Google Scholar]
  23. Techaarpornkul S., Barretto N., Peeples M. E. 2001; Functional analysis of recombinant respiratory syncytial virus deletion mutants lacking the small hydrophobic and/or attachment glycoprotein gene. J Virol 75:6825–6834 [CrossRef]
     [Google Scholar]
  24. Teng M. N., Whitehead S. S., Collins P. L. 2001; Contribution of the respiratory syncytial virus G glycoprotein and its secreted and membrane-bound forms to virus replication in vitro and in vivo . Virology 289:283–296 [CrossRef]
     [Google Scholar]
  25. Thiel V., Karl N., Schelle B., Disterer P., Klagge I., Siddell S. G. 2003; Multigene RNA vector based on coronavirus transcription. J Virol 77:9790–9798 [CrossRef]
     [Google Scholar]
  26. van den Hoogen B. G., de Jong J. C., Groen J., Kuiken T., de Groot R., Fouchier R. A. M., Osterhaus A. D. M. E. 2001; A newly discovered human pneumovirus isolated from young children with respiratory tract disease. Nat Med 7:719–724 [CrossRef]
     [Google Scholar]
  27. van den Hoogen B. G., Bestebroer T. M., Osterhaus A. D. M. E., Fouchier R. A. M. 2002; Analysis of the genomic sequence of a human metapneumovirus. Virology 295:119–132 [CrossRef]
     [Google Scholar]
  28. van den Hoogen B. G., van Doornum G. J. J., Fockens J. C., Cornelissen J. J., Beyer W. E. P., de Groot R., Osterhaus A. D. M. E., Fouchier R. A. M. 2003; Prevalence and clinical symptoms of human metapneumovirus infection in hospitalized patients. J Infect Dis 188:1571–1577 [CrossRef]
     [Google Scholar]
  29. Whitehead S. S., Bukreyev A., Teng M. N., Firestone C.-Y., St Claire M., Elkins W. R., Collins P. L., Murphy B. R. 1999; Recombinant respiratory syncytial virus bearing a deletion of either the NS2 or SH gene is attenuated in chimpanzees. J Virol 73:3438–3442
     [Google Scholar]
  30. Wilding G. P., Baxter-Jones C., Grant M. 1986; Ciliostatic agent found in rhinotracheitis. Vet Rec 118:735
     [Google Scholar]
  31. Yu Q., Davis P. J., Brown T. D. K., Cavanagh D. 1992; Sequence and in vitro expression of the M2 gene of turkey rhinotracheitis pneumovirus. J Gen Virol 73:1355–1363 [CrossRef]
     [Google Scholar]
  32. Yu Q., Barrett T., Brown T. D. K., Cook J. K. A., Green P., Skinner M. A., Cavanagh D. 1994; Protection against turkey rhinotracheitis pneumovirus (TRTV) induced by a fowlpox virus recombinant expressing the TRTV fusion glycoprotein (F). Vaccine 12:569–573 [CrossRef]
     [Google Scholar]
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Development of a reverse-genetics system for Avian pneumovirus demonstrates that the small hydrophobic (SH) and attachment (G) genes are not essential for virus viability
J. Gen. Virol. 85, 3219 (2004); https://doi.org/10.1099/vir.0.80229-0
/content/journal/jgv/10.1099/vir.0.80229-0
/content/journal/jgv/10.1099/vir.0.80229-0
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