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Molecular Cloning of the Nucleoprotein Gene of Canine Distemper Virus

Division of Virology, Department of Pathology, University of Cambridge, Laboratories Block, Addenbrooke's Hospital, Hills Road, Cambridge CB2 2QQ, U.K.

Messenger RNAs labelled in vivo in Vero cells infected with canine distemper virus were analysed by electrophoresis on 1.5% agarose gels containing 2M-formaldehyde. Seven virus-specific RNA bands could be distinguished which were not sensitive to actinomycin D treatment and were confined to the polyadenylated RNA fraction. The most abundant virus-specific mRNA species had a molecular weight of 0.52 x 10⁶ and its coding capacity was consistent with it being the mRNA for the most abundant virus-specific protein, the nucleoprotein. Polyadenylated RNA of this size class was purified by electrophoresis on a polyacrylamide gel and cloned into the PstI site of plasmid pBR322. A virus-specific clone obtained, clone 224, was then used to select messenger RNA from infected cells. The messenger RNA selected had a molecular weight of 0.52 x 10⁶ and directed the synthesis of only the virus-specific nucleoprotein when used to stimulate a wheat germ cell-free system.

Keywords: CDV, cloning, NP gene

Received 10 October 1983; accepted 2 December 1983.