This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by de Haan, C. A. M.
Right arrow Articles by Rottier, P. J. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by de Haan, C. A. M.
Right arrow Articles by Rottier, P. J. M.

 Previous Article  |  Next Article 

Journal of Virology, October 2005, p. 12742-12751, Vol. 79, No. 20
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.20.12742-12751.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Coronaviruses as Vectors: Stability of Foreign Gene Expression

Cornelis A. M. de Haan,* Bert Jan Haijema, David Boss, Frank W. H. Heuts, and Peter J. M. Rottier

Virology Division, Department of Infectious diseases and Immunology, Faculty of Veterinary Medicine and Institute of Biomembranes, Utrecht University, 3584 CL Utrecht, The Netherlands

Received 6 April 2005/ Accepted 1 August 2005

Coronaviruses are enveloped, positive-stranded RNA viruses considered to be promising vectors for vaccine development, as (i) genes can be deleted, resulting in attenuated viruses; (ii) their tropism can be modified by manipulation of their spike protein; and (iii) heterologous genes can be expressed by simply inserting them with appropriate coronaviral transcription signals into the genome. For any live vector, genetic stability is an essential requirement. However, little is known about the genetic stability of recombinant coronaviruses expressing foreign genes. In this study, the Renilla and the firefly luciferase genes were systematically analyzed for their stability after insertion at various genomic positions in the group 1 coronavirus feline infectious peritonitis virus and in the group 2 coronavirus mouse hepatitis virus. It appeared that the two genes exhibit intrinsic differences, the Renilla gene consistently being maintained more stably than the firefly gene. This difference was not caused by genome size restrictions, by different effects of the encoded proteins, or by different consequences of the synthesis of the additional subgenomic mRNAs. The loss of expression of the firefly luciferase was found to result from various, often large deletions of the gene, probably due to RNA recombination. The extent of this process appeared to depend strongly on the coronaviral genomic background, the luciferase gene being much more stable in the feline than in the mouse coronavirus genome. It also depended significantly on the particular genomic location at which the gene was inserted. The data indicate that foreign sequences are more stably maintained when replacing nonessential coronaviral genes.

* Corresponding author. Mailing address: Virology Division, Department of Infectious Diseases and Immunology, Yalelaan 1, 3584CL Utrecht, The Netherlands. Phone: 31-30-2534195. Fax: 31-30-2536723. E-mail: x.haan{at}vet.uu.nl.

Journal of Virology, October 2005, p. 12742-12751, Vol. 79, No. 20
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.20.12742-12751.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Gadlage, M. J., Graham, R. L., Denison, M. R. (2008). Murine Coronaviruses Encoding nsp2 at Different Genomic Loci Have Altered Replication, Protein Expression, and Localization. J. Virol. 82: 11964-11969 [Abstract] [Full Text]  
  • Zhang, X., Nuss, D. L. (2008). A host dicer is required for defective viral RNA production and recombinant virus vector RNA instability for a positive sense RNA virus. Proc. Natl. Acad. Sci. USA 105: 16749-16754 [Abstract] [Full Text]  
  • Tekes, G., Hofmann-Lehmann, R., Stallkamp, I., Thiel, V., Thiel, H.-J. (2008). Genome Organization and Reverse Genetic Analysis of a Type I Feline Coronavirus. J. Virol. 82: 1851-1859 [Abstract] [Full Text]  
  • van der Meer, F. J. U. M., de Haan, C. A. M., Schuurman, N. M. P., Haijema, B. J., Verheije, M. H., Bosch, B. J., Balzarini, J., Egberink, H. F. (2007). The carbohydrate-binding plant lectins and the non-peptidic antibiotic pradimicin A target the glycans of the coronavirus envelope glycoproteins. J Antimicrob Chemother 60: 741-749 [Abstract] [Full Text]  
  • Han, J., Liu, G., Wang, Y., Faaberg, K. S. (2007). Identification of Nonessential Regions of the nsp2 Replicase Protein of Porcine Reproductive and Respiratory Syndrome Virus Strain VR-2332 for Replication in Cell Culture. J. Virol. 81: 9878-9890 [Abstract] [Full Text]  
  • de Haan, C. A. M., te Lintelo, E., Li, Z., Raaben, M., Wurdinger, T., Bosch, B. J., Rottier, P. J. M. (2006). Cooperative Involvement of the S1 and S2 Subunits of the Murine Coronavirus Spike Protein in Receptor Binding and Extended Host Range. J. Virol. 80: 10909-10918 [Abstract] [Full Text]  
  • Wurdinger, T., Verheije, M. H., Broen, K., Bosch, B. J., Haijema, B. J., de Haan, C. A. M., van Beusechem, V. W., Gerritsen, W. R., Rottier, P. J. M. (2005). Soluble Receptor-Mediated Targeting of Mouse Hepatitis Coronavirus to the Human Epidermal Growth Factor Receptor. J. Virol. 79: 15314-15322 [Abstract] [Full Text]  
  • de Haan, C. A. M., Li, Z., te Lintelo, E., Bosch, B. J., Haijema, B. J., Rottier, P. J. M. (2005). Murine Coronavirus with an Extended Host Range Uses Heparan Sulfate as an Entry Receptor. J. Virol. 79: 14451-14456 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»