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Journal of Virology, April 2006, p. 3670-3674, Vol. 80, No. 7
0022-538X/06/$08.00+0     doi:10.1128/JVI.80.7.3670-3674.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Recovery of a Neurovirulent Human Coronavirus OC43 from an Infectious cDNA Clone{dagger}

Julien R. St-Jean,1 Marc Desforges,1 Fernando Almazán,2 Hélène Jacomy,1 Luis Enjuanes,2 and Pierre J. Talbot1*

Laboratory of Neuroimmunovirology, INRS-Institut Armand-Frappier, 531 Boulevard des Prairies, Laval, Québec H7V 1B7, Canada,1 Department of Molecular and Cell Biology, Centro Nacional de Biotecnología, CSIC, Campus Universidad Autónoma, Darwin 3, 28049 Madrid, Spain2

Received 23 October 2005/ Accepted 9 January 2006

This study describes the assembly of a full-length cDNA clone of human coronavirus (HCoV)-OC43 in a bacterial artificial chromosome (BAC). The BAC containing the full-length infectious cDNA (pBAC-OC43FL) was assembled using a two-part strategy. The first step consisted in the introduction of each end of the viral genome into the BAC with accessory sequences allowing proper transcription. The second step consisted in the insertion of the whole HCoV-OC43 cDNA genome into the BAC. To produce recombinant viral particles, pBAC-OC43FL was transfected into BHK-21 cells. Recombinant virus displayed the same phenotypic properties as the wild-type virus, including infectious virus titers produced in cell culture and neurovirulence in mice.

* Corresponding author. Mailing address: Laboratory of Neuroimmunovirology, INRS-Institut Armand-Frappier, 531 Boulevard des Prairies, Laval H7V 1B7, Québec, Canada. Phone: (450) 686-5515. Fax: (450) 686-5566. E-mail: pierre.talbot{at}iaf.inrs.ca.

{dagger} Supplemental material for this article may be found at http://jvi.asm.org/.

Journal of Virology, April 2006, p. 3670-3674, Vol. 80, No. 7
0022-538X/06/$08.00+0     doi:10.1128/JVI.80.7.3670-3674.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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