SUMMARYThe complete nucleotide sequence of the RNA of an aphid non-transmissible plum pox virus (PPV-NAT) isolate has been determined from five overlapping cDNA clones, cDNA prepared by primer extension was used to determine the 5' terminus. The assembled RNA is 9741 nucleotides in length, excluding a 3' terminal poly(A) sequence. One large open reading frame starts at nucleotide positions 36 to 38 and is terminated with an UAG codon at positions 9522 to 9524. The putative start codon is located at positions 147 to 149. The encoded polyprotein has a predicted Mr of 353"8K. Comparison of cistrons from tobacco vein mottling virus and tobacco etch virus with those predicted for PPV-NAT indicated a similar genome organization. A highly conserved sequence of 12 nucleotides was found in the 5' non-coding region of these three potyviruses. The potential polyadenylation signal from yeast (UAUGU) was found in the 3' non-coding region of PPV-NAT and several other members of the potyvirus group.
Genomic RNA of plum pox virus (PPV) was used as a template for the synthesis of complementary DNA (cDNA). The generated cDNA molecules were subsequently cloned into pBR 322. A physical map covering 9700 bases of the PPV genome was constructed from 8, clones by hybridization and restriction endonuclease digestion. Clone pPPV‐NAT 309, starting at the 3′‐end, with an 866 bp insert was used in Northern‐ and Dot‐hybridizations for the detection of single‐stranded viral RNA in total nucleic acid as well as in sap preparations of PPV infected Nicotiana clevelandii. The nucleotide sequence of this clone was determined, the amino acid sequence of the coat protein C‐terminal part was deduced and compared with four other coat proteins of potyviruses.
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