Genome organization and reverse genetic analysis of a type I feline coronavirus.
In this study we report the complete sequence and genome organization of the serotype I feline coronavirus (FCoV) strain Black. Furthermore, a reverse genetic system was established for this FCoV strain by cloning a full-length cDNA copy into vaccinia virus. This clone served as basis for the generation of recombinant FCoV (recFCoV) that was shown to bear the same features in vitro as the parental FCoV. Using this system, accessory 3abc genes in the FCoV genome were replaced by green fluorescent protein (recFCoV-GFP) and Renilla luciferase genes (recFCoV-RL). In addition, we showed that feline CD14(+) blood monocytes and dendritic cells can be easily detected after infection with recFCoV-GFP. Thus, our established reverse genetic system provides a suitable tool to study the molecular biology of serotype I FCoV.
Tekes
Gergely
G
Kantonal Hospital St. Gallen, Research Department, 9007 St. Gallen, Switzerland.
Hofmann-Lehmann
Regina
R
Stallkamp
Iris
I
Thiel
Volker
V
Thiel
Heinz-Jürgen
HJ
D000818:Animals; D001483:Base Sequence; D002415:Cats; D003001:Cloning, Molecular; D016765:Coronavirus, Feline / Q000235:genetics*; D018076:DNA, Complementary / Q000235:genetics; D003713:Dendritic Cells / Q000276:immunology; D023061:Gene Order; D017930:Genes, Reporter; D016679:Genome, Viral; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D018950:Lipopolysaccharide Receptors / Q000032:analysis; D049410:Luciferases, Renilla / Q000032:analysis / Q000235:genetics; D008969:Molecular Sequence Data; D009000:Monocytes / Q000276:immunology; D017422:Sequence Analysis, DNA / Q000379:methods*; D012703:Serotyping
D018076:DNA, Complementary; D018950:Lipopolysaccharide Receptors; D049452:Green Fluorescent Proteins; D049410:Luciferases, Renilla