cDNA Cloning of the Messenger RNAs of Five Genes of Canine Distemper Virus

Abstract

SUMMARYMessenger RNAs from Vero cells infected with the Onderstepoort strain of canine distemper virus (CDV) were cloned into the PstI site of plasmid pATI53. Total polyadenylated RNA was used and resulting clones were screened with 3zp-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.

Some Highlights of Animal Virus Research in 1985

“…Gene order appears to be similar to that of Sendal virus, with the arrangement of the distal genes (in the 5' portion of genome RNA) remaining tentative Rozenblatt et al, 1985;Russell et al, 1985). Most interestingly, both viruses possess a second reading frame in the P transcript, like Sendal virus, and this is probably translated from the same mRNA species as P (Barrett et aL, 1985 ;.…”

Characterization of Clones for the Sixth (L) Gene and a Transcriptional Map for Morbilliviruses

“…Furthermore, in CDVinfected cells readthrough RNAs representing three tandem genes, N -P -M and P -M -F , were identified and in MV-infected cells a probable M -F -H readthrough product was observed. These data and those obtained previously by Richardson et al (1985) and Russell et al (1985) allowed the construction of a complete transcription map for the morbilliviruses, shown in diagrammatic form in Fig. 4.…”

“…As outlined in the Introduction, partial or complete cDNA clones of five mRNAs of CDV and MV have been described by ourselves and other groups and the mRNA profiles induced in MVand CDV-infected cells have been characterized (Hall et al, 1978;Udem & Cook, 1984;Yoshikawa & Yamanouchi, 1984;Barrett & Underwood, 1985;Russell et al, 1985). Throughout this paper we refer to the numbering of mRNA species used in the report by Barrett & Underwood (1985).…”

“…The following CDV clones described by Russell et al (1985) were used: pCDV51 (M), pCDV102 (N), pCDV2 (F), pCDV28 (N), pCDV30 (P), pCDV22 (P), pCDV54 (H) and pCDV65 (L) (this paper). MVspecific clones used were clone 15 ~ CL-N (Gorecki & Rozenblatt, 1980), CL-P , CL-M (Rozenblatt et al, 1982), clones 3 (F/H) and 2 (H) (Billeter et al, 1984;Baczko et al, 1986), pMV 14 (H) (J. D. Hull & B. K. Rima, unpublished results) and clone 6 (L) (Billeter et al, 1984;this paper) and MV 03 (Billeter et al, 1984;Rozenblatt et al, 1985a).…”

“…cDNA cloning and base sequencing of the N genes of MV (Gorecki & Rozenblatt, t 980;Billeter et al, 1984; and of CDV (Barrett & Mahy, 1984;Russell et al, 1985 ;Rozenblatt et al, 1985b), the P (C) genes of MV and of CDV Russell et al, 1985;Rozenblatt et al, 1985b) and the M proteins of MV (Rozenblatt et al, 1982;Bellini et al, 1985) and of CDV (Rozenblatt et al, 1985b;Russell et al, 1985;Bellini et al, 1986) have been described. Clones of the mRNAs coding presumably for the two glycoproteins of these viruses have been described for CDV and MV (Billeter et al, 1984).…”