1887

Journal of General Virology header logo

Volume 81, Issue 7

Expression of reporter genes from the defective RNA CD-61 of the coronavirus infectious bronchitis virus Free

Abstract

The defective RNA (D-RNA) CD-61, derived from the Beaudette strain of the avian coronavirus infectious bronchitis virus (IBV), was used as an RNA vector for the expression of two reporter genes, luciferase and chloramphenicol acetyltransferase (CAT). D-RNAs expressing the CAT gene were demonstrated to be capable of producing CAT protein in a helper-dependent expression system to about 1·6 μg per 10 cells. The reporter genes were expressed from two different sites within the CD-61 sequence and expression was not affected by interruption of the CD-61-specific ORF. Expression of the reporter genes was under the control of a transcription-associated sequence (TAS) derived from the Beaudette gene 5, normally used for the transcription of IBV subgenomic mRNA 5. The Beaudette gene 5 TAS is composed of two tandem repeats of the IBV canonical consensus sequence involved in the acquisition of a leader sequence during the discontinuous transcription of IBV subgenomic mRNAs. It is demonstrated that only one canonical sequence is required for expression of mRNA 5 or for the expression of an mRNA from a D-RNA and that either sequence can function as an acceptor site for acquisition of the leader sequence.

  • Received:
  • Accepted:
  • Published Online:
© Microbiology Society
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-81-7-1687
2000-07-01
2024-04-20
Loading full text...

Full text loading...

/deliver/fulltext/jgv/81/7/0811687a.html?itemId=/content/journal/jgv/10.1099/0022-1317-81-7-1687&mimeType=html&fmt=ahah

References

  1. Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A. & Struhl, K. (editors) (1987).Current Protocols in Molecular Biology. New York: John Wiley.
  2. Baric, R. S., Stohlman, S. A. & Lai, M. M. C. (1983). Characterization of replicative intermediate RNA of mouse hepatitis virus: presence of leader RNA sequences on nascent chains.Journal of Virology 48, 633-640. [Google Scholar]
  3. Binns, M. M., Boursnell, M. E. G., Cavanagh, D., Pappin, D. J. C. & Brown, T. D. K. (1985). Cloning and sequencing of the gene encoding the spike protein of the coronavirus IBV.Journal of General Virology 66, 719-726.[CrossRef] [Google Scholar]
  4. Binns, M. M., Boursnell, M. E. G., Tomley, F. M. & Brown, T. D. K. (1986). Comparison of the spike precursor sequences of coronavirus IBV strains M41 and 6/82 with that of IBV Beaudette.Journal of General Virology 67, 2825-2831.[CrossRef] [Google Scholar]
  5. Boursnell, M. E. G., Brown, T. D. K., Foulds, I. J., Green, P. F., Tomley, F. M. & Binns, M. M. (1987). Completion of the sequence of the genome of the coronavirus avian infectious bronchitis virus.Journal of General Virology 68, 57-77.[CrossRef] [Google Scholar]
  6. Cavanagh, D. & Naqi, S. (1997). Infectious bronchitis. In Diseases of Poultry, pp. 511-526. Edited by B. W. Calnek, H. J. Barnes, C. W. Beard, W. M. Reid & H. W. Yoda. Ames, IA: Iowa State University Press.
  7. Darbyshire, J. H., Rowell, J. G., Cook, J. K. A. & Peters, R. W. (1979). Taxonomic studies on strains of avian infectious bronchitis virus using neutralisation tests in tracheal organ cultures.Archives of Virology 61, 227-238.[CrossRef] [Google Scholar]
  8. Davelaar, F. G., Kouwenhoven, B. & Burger, A. G. (1984). Occurrence and significance of infectious bronchitis virus variant strains in egg and broiler production in the Netherlands.Veterinary Quarterly 6, 114-120.[CrossRef] [Google Scholar]
  9. Feinberg, A. P. & Vogelstein, B. (1983). A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity.Analytical Biochemistry 132, 6-13.[CrossRef] [Google Scholar]
  10. Hiscox, J. A., Mawditt, K. L., Cavanagh, D. & Britton, P. (1995). Investigation of the control of coronavirus subgenomic mRNA transcription by using T7-generated negative-sense RNA transcripts.Journal of Virology 69, 6219-6227. [Google Scholar]
  11. Izeta, A., Smerdou, C., Alonso, S., Pénzes, Z., Mendez, A., Plana-Durán, J. & Enjuanes, L. (1999). Replication and packaging of transmissible gastroenteritis coronavirus-derived synthetic minigenomes.Journal of Virology 73, 1535-1545. [Google Scholar]
  12. Jia, W. & Naqi, S. A. (1997). Sequence analysis of gene 3, gene 4 and gene 5 of avian infectious bronchitis virus strain CU-T2.Gene 189, 189-193.[CrossRef] [Google Scholar]
  13. Joo, M. & Makino, S. (1992). Mutagenic analysis of the coronavirus intergenic consensus sequence.Journal of Virology 66, 6330-6337. [Google Scholar]
  14. Joo, M. & Makino, S. (1995). The effect of two closely inserted transcription consensus sequences on coronavirus transcription.Journal of Virology 69, 272-280. [Google Scholar]
  15. Krishnan, R., Chang, R. Y. & Brian, D. A. (1996). Tandem placement of a coronavirus promoter results in enhanced mRNA synthesis from the downstream-most initiation site.Virology 218, 400-405.[CrossRef] [Google Scholar]
  16. Kusters, J. G., Niesters, H. G. M., Lenstra, J. A., Horzinek, M. C. & van der Zeijst, B. A. M. (1989). Phylogeny of antigenic variants of avian coronavirus IBV. Virology 169, 217-221.[CrossRef] [Google Scholar]
  17. Lai, M. M., Zhang, X., Hinton, D. & Stohlman, S. (1997). Modulation of mouse hepatitis virus infection by defective-interfering RNA-mediated expression of viral proteins and cytokines.Journal of Neurovirology 3, S33-S34. [Google Scholar]
  18. Lambrechts, C., Pensaert, M. & Ducatelle, R. (1993). Challenge experiments to evaluate cross-protection induced at the trachea and kidney level by vaccine strains and Belgian nephropathogenic isolates of avian infectious bronchitis virus.Avian Pathology 22, 577-590.[CrossRef] [Google Scholar]
  19. Liao, C.-L. & Lai, M. M. C. (1994). Requirement of the 5′-end genomic sequence as an upstream cis-acting element for coronavirus subgenomic mRNA transcription.Journal of Virology 68, 4727-4737. [Google Scholar]
  20. Liao, C.-L., Zhang, X. & Lai, M. M. C. (1995). Coronavirus defective-interfering RNA as an expression vector: the generation of a pseudorecombinant mouse hepatitis virus expressing hemagglutinin–esterase.Virology 208, 319-327.[CrossRef] [Google Scholar]
  21. Makino, S. & Joo, M. (1993). Effect of intergenic consensus sequence flanking sequences on coronavirus transcription.Journal of Virology 67, 3304-3311. [Google Scholar]
  22. Makino, S. & Lai, M. M. C. (1989). High-frequency leader sequence switching during coronavirus defective interfering RNA replication.Journal of Virology 63, 5285-5292. [Google Scholar]
  23. Makino, S., Joo, M. & Makino, J. K. (1991). A system for study of coronavirus mRNA synthesis: a regulated, expressed subgenomic defective interfering RNA results from intergenic site insertion. Journal of Virology 65, 6031-6041. [Google Scholar]
  24. Meulemans, G., Carlier, M. C., Gonze, M., Petit, P. & Vandenbroeck, M. (1987). Incidence, characterisation and prophylaxis of nephropathogenic avian infectious bronchitis viruses. Veterinary Record 120, 205-206.[CrossRef] [Google Scholar]
  25. Niesters, H. G. M., Lenstra, J. A., Spaan, W. J. M., Zijderveld, A. J., Bleumink-Pluym, N. M. C., Hong, F., van Scharrenburg, G. J. M., Horzinek, M. C. & van der Zeijst, B. A. M. (1986). The peplomer protein sequence of the M41 strain of coronavirus IBV and its comparison with Beaudette strains.Virus Research 5, 253-263.[CrossRef] [Google Scholar]
  26. Pause, A., Belsham, G. J., Gingras, A. C., Donze, O., Lin, T. A., Lawrence, J. C.Jr & Sonenberg, N. (1994). Insulin-dependent stimulation of protein synthesis by phosphorylation of a regulator of 5′-cap function.Nature 371, 762-767.[CrossRef] [Google Scholar]
  27. Pénzes, Z., Tibbles, K., Shaw, K., Britton, P., Brown, T. D. K. & Cavanagh, D. (1994). Characterization of a replicating and packaged defective RNA of avian coronavirus infectious bronchitis virus.Virology 203, 286-293.[CrossRef] [Google Scholar]
  28. Pénzes, Z., Wroe, C., Brown, T. D., Britton, P. & Cavanagh, D. (1996). Replication and packaging of coronavirus infectious bronchitis virus defective RNAs lacking a long open reading frame.Journal of Virology 70, 8660-8668. [Google Scholar]
  29. Sambrook, J., Fritsch, E. F. & Maniatis, T. (1989).Molecular Cloning. A Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  30. Sawicki, S. G. & Sawicki, D. L. (1990). Coronavirus transcription: subgenomic mouse hepatitis virus replicative intermediates function in RNA synthesis.Journal of Virology 64, 1050-1056. [Google Scholar]
  31. Sawicki, S. G. & Sawicki, D. L. (1998). A new model for coronavirus transcription.Advances in Experimental Medicine and Biology 440, 215-219. [Google Scholar]
  32. Shaw, K., Britton, P. & Cavanagh, D. (1996). Sequence of the spike protein of the Belgian B1648 isolate of nephropathogenic infectious bronchitis virus.Avian Pathology 25, 607-611.[CrossRef] [Google Scholar]
  33. Stern, D. F. & Kennedy, S. I. T. (1980). Coronavirus multiplication strategy. I. Identification and characterization of virus-specific RNA.Journal of Virology 34, 665-674. [Google Scholar]
  34. Stirrups, K., Shaw, K., Evans, S., Dalton, K., Cavanagh, D. & Britton, P. (2000). Leader switching occurs during the rescue of defective RNAs by heterologous strains of the coronavirus infectious bronchitis virus.Journal of General Virology 81, 791-801. [Google Scholar]
  35. Sutou, S., Sato, S., Okabe, T., Nakai, M. & Sasaki, N. (1988). Cloning and sequencing of genes encoding structural proteins of avian infectious bronchitis virus.Virology 165, 589-595.[CrossRef] [Google Scholar]
  36. van der Most, R. G., de Groot, R. J. & Spaan, W. J. M. (1994). Subgenomic RNA synthesis directed by a synthetic defective interfering RNA of mouse hepatitis virus: a study of coronavirus transcription initiation.Journal of Virology 68, 3656-3666. [Google Scholar]
  37. van Marle, G., Luytjes, W., van der Most, R. G., van der Straaten, T. & Spaan, W. J. (1995). Regulation of coronavirus mRNA transcription.Journal of Virology 69, 7851-7856. [Google Scholar]
  38. Zhang, X., Hinton, D. R., Cua, D. J., Stohlman, S. A. & Lai, M. M. (1997). Expression of interferon-gamma by a coronavirus defective-interfering RNA vector and its effect on viral replication, spread, and pathogenicity.Virology 233, 327-338.[CrossRef] [Google Scholar]
  39. Zhang, X., Hinton, D. R., Park, S., Parra, B., Liao, C.-L., Lai, M. M. & Stohlman, S. A. (1998). Expression of hemagglutinin/esterase by a mouse hepatitis virus coronavirus defective-interfering RNA alters viral pathogenesis.Virology 242, 170-183.[CrossRef] [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-81-7-1687
Loading
Expression of reporter genes from the defective RNA CD-61 of the coronavirus infectious bronchitis virus
J. Gen. Virol. 81, 1687 (2000); https://doi.org/10.1099/0022-1317-81-7-1687
/content/journal/jgv/10.1099/0022-1317-81-7-1687
/content/journal/jgv/10.1099/0022-1317-81-7-1687
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error