Abstract
The large size of the Coronavirus genome has made reverse genetics difficult. Targeted recombination, a technique developed by Masters and colleagues, has facilitated the introduction of mutations into the Coronavirus genome (Kuo et al., 2000). Previous work by Skinner and Siddell showed that MHV-JHM ORF 4 encodes a 15 kDa protein composed of 139 amino acids. This protein is relatively rich in threonines and includes a hydrophobic region. The N-terminus contains a potential membrane-anchoring region and the C-terminus has a possible RNA binding region (Skinner and Siddell, 1985). MHV-S, a natural variant, does not encode a functional ORF 4 suggesting that the ORF 4 product was not necessary for growth in tissue culture cells or animals. Additionally this strain contained a deletion within ORF 5a (Yokomori and Lai, 1991). Lack of mRNA 4 synthesis most likely resulted from a point mutation in the intergenic sequence (UCUAAAC to UUUAAAC). In this study, targeted recombination was used to genetically disrupt ORF 4. This recombinant virus was then analyzed in a murine model of encephalitis.
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
Kuo, L., Godeke, G. J., Raamsman, M. J., Masters, P. S. and P. J. Rottier. 2000. Retargeting of Coronavirus by substitution of the spike glycoprotein ectodomain: crossing the host cell species barrier. J. Virol. 74(3): 1393–1406.
Perlman, S. Jacobsen, G., Olson, A. L., and A. Afifi. 1990. Identification of the spinal cord as a major site of persistence during chronic infection with a murine Coronavirus. Virology. 175:418–426.
Pewe, L., Wu, G. F., Barnett, E. M., Castro, R. F. and S. Perlman. 1996. Cytotoxic T cell-resistant variants are selected in a virus-induced demyelinating disease. Immunity. 5:253–262.
Reed, L. J. and H. Muench. 1938. A simple method of estimating fifty percent points. Am. J. Hygiene. 27:493–497.
Skinner, M. A. and S. G. Siddell. 1985. coding sequence of Coronavirus MHV-JHM mRNA 4. J. Gen. Virol. 66:593–596.
Weiss, S. R., Zoltick, P. W., and J. L. Leibowitz. 1993. The ns 4 gene of mouse hepatitis virus (MHV), strain A 59 contains two ORFs and thus differs from ns 4 of the JHM and S strains. Arch. Virol. 129:301–309.
Yokomori, K., and M. M. C. Lai. 1991. Mouse hepatitis virus S RNA sequence reveals that nonstructural proteins ns4 and ns5a are not essential for murine Coronavirus replication. J. Virol. 65(10):5605–5608.
Yu, X., Bi, W., Weiss, S. R. and J. L. Leibowitz. 1994. Mouse hepatitis virus gene 5b protein is a new virion envelope protein. Virology. 202:1018–1023.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2001 Springer Science+Business Media New York
About this chapter
Cite this chapter
Ontiveros, E., Kuo, L., Masters, P., Perlman, S. (2001). Analysis of Nonessential Gene Function in Recombinant MHV-JHM. In: Lavi, E., Weiss, S.R., Hingley, S.T. (eds) The Nidoviruses. Advances in Experimental Medicine and Biology, vol 494. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-1325-4_13
Download citation
DOI: https://doi.org/10.1007/978-1-4615-1325-4_13
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4613-5498-7
Online ISBN: 978-1-4615-1325-4
eBook Packages: Springer Book Archive