The translation products directed by cucumber necrosis virus (CNV) RNA were analysed in both rabbit reticulocyte lysate and wheatgerm extract cell-free translation systems. In rabbit reticulocyte lysates, one major protein of approximate 34.6K was produced. In wheatgerm extracts, four proteins of approximate values 41.6K, 34.6K, 24K and 20K were produced. The genomic locations of the CNV translation products were determined using several experimental approaches including, first, hybrid-arrested translation using negative-sense RNA corresponding to selected regions of the CNV genome, second, translation of synthetic positive-sense CNV transcripts and third, translation of CNV virion RNA fractionated according to size. Together these experiments demonstrated that the protein of 34.6K is derived from the 5′-proximal coding region, the 41.6K protein is derived from an internal coding region, and that at least one but probably both the 24K and 20K proteins are derived from the 3′-terminal coding region. In addition, immunoprecipitation of translation products using anti-CNV polyclonal serum demonstrated that the 41.6K protein is the coat protein. The templates for the expression of CNV cistrons were investigated by translation of sucrose gradient-fractionated CNV virion RNA as well as translation of positive-sense synthetic transcripts.


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