Recovery of a neurovirulent human coronavirus OC43 from an infectious cDNA clone.
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-OC43(FL)) 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-OC43(FL) 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.
Laboratory of Neuroimmunovirology, INRS-Institut Armand-Frappier, 531 Boulevard des Prairies, Laval H7V 1B7, Québec, Canada.
Julien R
JR
St-Jean
Marc
M
Desforges
Fernando
F
Almazán
Hélène
H
Jacomy
Luis
L
Enjuanes
Pierre J
PJ
Talbot
D000818:Animals; D002460:Cell Line; D020805:Central Nervous System Viral Diseases / Q000503:physiopathology* / Q000821:virology; D022202:Chromosomes, Artificial, Bacterial; D002999:Clone Cells; D003001:Cloning, Molecular; D018352:Coronavirus Infections / Q000503:physiopathology / Q000821:virology; D028962:Coronavirus OC43, Human / Q000235:genetics / Q000472:pathogenicity / Q000502:physiology; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics*; D016679:Genome, Viral; D006801:Humans; D007276:Injections, Intraventricular; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D009417:Nerve Tissue / Q000821:virology*; D011995:Recombination, Genetic; D015996:Survival Rate; D014162:Transfection; D014771:Virion; D014774:Virulence; D019065:Virus Assembly; D014779:Virus Replication
D018076:DNA, Complementary