Abstract
The Beaudette strain of IBV was passaged 16 times in chick kidney (CK) cells. Total cellular RNA was analyzed by Northern hybridization and was probed with 32P-labeled cDNA probes corresponding to the first 2 kb of the 5′ end of the genome, but excluding the leader, and to the last 1.8 kb of the 3′ end of the genome. A new, defective IBV RNA species (CD-91) was detected at passage six. The defective RNA, present in total cell extract RNA and in oligo-(dT)30-selected RNA from passage 15, was amplified by the reverse transcription-polymerase chain reaction (RT-PCR) to give four fragments. The oligonucleotides used were selected such that CD-91 RNA, but not the genomic RNA, would be amplified. Cloning and sequencing of the PCR products showed that CD-91 comprises 9.1 kb and has three regions of the genome. It contains 1133 nucleotides from the 5′ end of the genome, 6322 from gene lb corresponding to position 12423 to 18744 in the IBV genome and 1626 from the 3’ end of the genome. At position 749 one nucleotide, an adenine residue, was absent from CD-91 RNA. By Northern hybridization CD-91 RNA was detected in virions in higher amounts than the subgenomic mRNAs.
Sequence data from this article have been deposited with the EMBL, GenBank and DDJB Nucleotide Sequence Databases under Accession No. Z30541
Chapter PDF
Similar content being viewed by others
Keywords
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.
References
Baric, R. S., Stohlman, S. A., Razavi, M. K., and Lai, M. M. C. (1985). Characterisation of leader related small RNAs in coronavirus infected cells. Further evidence for leader primed mechanism transcription. Virus Res. 3, 19–33.
Baric, R. S., Shieh, C., Stohlman, S. A., and Lai, M. M. C. (1987). Analysis of intracellular small RNAs of mouse hepatitis virus: evidence for discontinuous transcription. Virology 156, 342–354.
Boursnell, M. E. G., Brown, T. D. K., Foulds, I. J., Green, P. F., Tomley, F. M., and Binns, M. M. (1987). Completion of the sequence of the genome of the coronavirus avian infectious bronchitis virus. J. Gen. Virol. 68, 57–77.
Cavanagh, D., Davis, P. J., and Cook, J. K. A. (1992). Infectious bronchitis virus: evidence for recombination within the Massachusetts serotype. Avian Pathol. 21,401–408.
de Groot, R. J., van der Most, R. G., and Spaan, W. J. M. (1992). The fitness of defective interfering murine coronavirus DI-a and its derivatives is decreased by nonsense and frameshift mutations. J. Virol.66, 5898–5905.
Fosmire, J. A., Hwang, K., and Makino, S. (1992). Identification and characterization of a coronavirus packaging signal. J. Virol. 66, 3522–3530.
Furuya, T., Macnaughton, T. B., La Monica, N., and Lai, M. M. C. (1993). Natural evolution of coronavirus defective-interfering RNA involves RNA recombination. Virology 194, 408–413.
Jeong, Y. S., and Makino, S. (1994). Evidence for coronavirus discontinuous transcription. J. Virol. 68, 2615–2623.
Kim, Y., Jeong, Y., and Makino, S. (1993). Analysis of cis-acting sequences essential for coronavirus defective interfering RNA replication. Virology 197, 53–63.
Lai, M. M. C., Baric, R. S., Makino, S., Keck, J. G., Egbert, J., Leibowitz, J. L., and Stohlman, S. A. (1985). Recombination between nonsegmented RNA genomes of murine coronaviruses. J. Virol. 56,449–456.
Lin, Y., and Lai, M. M. C. (1993). Deletion mapping of a mouse hepatitis virus defective interfering RNA reveals the requirement of an internal and discontiguous sequence for replication. J. Virol. 67,6110–6118.
Makino, S., Taguchi, F., and Fujiwara, K. (1984). Defective interfering particles of mouse hepatitis virus.Virology 133, 9–17.
Makino, S., Fujioka, N., and Fujiwara, K. (1985). Structure of the intracellular viral RNAs of defective interfering particles of mouse hepatitis virus. J. Virol 54, 329–336.
Makino, S., Shieh, C., Soe, L., Baker, S. C, and Lai, M. M. C. (1988). Primary structure and translation of a defective interfering RNA of murine coronavirus. Virology 166, 550–560.
Makino, S., Yokomori, K., and Lai, M. M. C. (1990). Analysis of efficiently packaged defective interfering RNAs of murine coronavirus: localization of a possible RNA-packaging signal. J. Virol 64, 6045–6053.
Sethna, P. B., Hofmann, M. A., and Brian, D. A. (1991). Minus-strand copies of replicating coronavirus mRNAs contain antileaders. J. Virol 65, 320–325.
van der Most, R. G., Bredenbeek, P. J., and Spaan, W. J. M. (1991). A domain at the 3’ end of the polymerase gene is essential for encapsidation of coronavirus defective interfering RNAs. J. Virol 65,3219–3226.
Zhao, X., Shaw, K., and Cavanagh, D. (1993). Presence of subgenomic mRNAs in virions of coronavirus IBV. Virology 196, 172–178.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1995 Springer Science+Business Media New York
About this chapter
Cite this chapter
Penzes, Z., Tibbles, K.W., Shaw, K., Britton, P., Brown, T.D.K., Cavanagh, D. (1995). Generation of a Defective RNA of Avian Coronavirus Infectious Bronchitis Virus (IBV). In: Talbot, P.J., Levy, G.A. (eds) Corona- and Related Viruses. Advances in Experimental Medicine and Biology, vol 380. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-1899-0_90
Download citation
DOI: https://doi.org/10.1007/978-1-4615-1899-0_90
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4613-5775-9
Online ISBN: 978-1-4615-1899-0
eBook Packages: Springer Book Archive