Expression in transgenic plants of a viral gene product that mediates insect transmission of potyviruses.

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.

[1]  R. Rhoads,et al.  Infectious in vitro transcripts from cloned cDNA of a potyvirus, tobacco vein mottling virus. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[2]  J. Carrington,et al.  A second proteinase encoded by a plant potyvirus genome. , 1989, The EMBO journal.

[3]  R. Rhoads,et al.  In vitro analysis of tobacco vein mottling virus NIa cistron: evidence for a virus-encoded protease. , 1988, Virology.

[4]  D. Hildebrand,et al.  Design and construction of a versatile system for the expression of foreign genes in plants. , 1987, Gene.

[5]  Robert E. Rhoads,et al.  The nucleotide sequence of tobacco vein mottling virus RNA , 1986, Nucleic Acids Res..

[6]  Pirone Tp,et al.  The involvement of a helper component in nonpersistent transmission of plant viruses by aphids. , 1984 .

[7]  M. Kozak Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs. , 1984, Nucleic acids research.

[8]  M. Van Montagu,et al.  Ti plasmid vector for the introduction of DNA into plant cells without alteration of their normal regeneration capacity , 1983, The EMBO journal.

[9]  A. Feinberg,et al.  A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. , 1983, Analytical biochemistry.

[10]  D. Ward,et al.  Rapid and sensitive colorimetric method for visualizing biotin-labeled DNA probes hybridized to DNA or RNA immobilized on nitrocellulose: Bio-blots. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[11]  D. Hanahan Studies on transformation of Escherichia coli with plasmids. , 1983, Journal of molecular biology.

[12]  W. Rutter,et al.  Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. , 1979, Biochemistry.

[13]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[14]  D. Govier,et al.  A virus-induced component of plant sap needed when aphids acquire potato virus Y from purified preparations. , 1974, Virology.