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  • Title: Expression in transgenic plants of a viral gene product that mediates insect transmission of potyviruses.
    Author: Berger PH, Hunt AG, Domier LL, Hellmann GM, Stram Y, Thornbury DW, Pirone TP.
    Journal: Proc Natl Acad Sci U S A; 1989 Nov; 86(21):8402-6. PubMed ID: 2813397.
    Abstract:
    Helper component (HC) is a virus-encoded nonstructural protein that is required for transmission of potyviruses by their aphid vectors. As a prelude to studies on the molecular basis of HC activity, a cDNA clone (pPB-3) was constructed that contained the first three cistrons (34 kDa-HC-42 kDa) of the RNA genome of the potyvirus tobacco vein mottling virus, the first six nucleotides of the adjacent cylindrical inclusion body protein cistron, and a synthetic translation termination codon. This construction was introduced into tobacco cells via a Ti plasmid-based vector. Northern blot analysis of transgenic plants demonstrated the presence of an RNA of the size expected from the construction of pPB-3, and Western blot analysis revealed the presence of a protein that comigrated with authentic HC, indicating that the proteolytic activity necessary to produce mature-sized HC was encoded by pPB-3. The HC produced in the transgenic plants was demonstrated to be active in a virus transmission bioassay with aphids.
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