cDNA cloning of the messenger RNAs of five genes of canine distemper virus

Abstract

Messenger RNAs from Vero cells infected with the Onderstepoort strain of canine distemper virus (CDV) were cloned into the PstI site of plasmid pAT153. Total polyadenylated RNA was used and resulting clones were screened with 32P-labelled cDNA probes from infected and mock-infected cells. The virus specificity of the clones was proven by Northern blot hybridization and by ability to select radioactive virus mRNAs labelled in vivo in the presence of actinomycin D. Clones from the N, P and M genes of CDV were identified by hybrid select translation; clones which presumably represent the H and F genes were also obtained. The clones allowed a designation of the major viral mRNA bands. Bicistronic mRNAs were identified, and their selection by various clones suggests a gene order of 3'-N-P-M-70K-65K-L-5' for this virus.