Plant expression vector pBI 121 containing the gene encoding coat protein of Plum Pox Virus of the Skierniewice isolate (CP PPV-S) was prepared (clone pCMl). The construct was used for transformation of Nicotiana tabacum plants using an Agrobacterium tumefaciens based system. About 82% of kanamycin resistant plant lines contained a transgene (the sequence of CP PPV-S) but only 81% of them actively expressed the PPV-S coat protein gene as measured by RT-PCR.
1. I.indbo, J.A., Silva-Rosales, L. & Dougherty, W.G. (1993) Pathogen derived resistance to potyviruses: working, but why? Setninars in Virology 4,369-379.
2. Fang, G. & Grumet, R. (1993) Genetic engineering of potyvirus resistance using constructs derived from the zucchini yellow mosaic virus coat protein gene. Molecular Plant-Microbe Interactions 6,358-367.
3. Reimann-Philipp, U. & Beachy, R.N. (1993) Coat protein-mediated resistance in transgenic tobacco expressing the tobacco mosaic virus coat protein from tissue-specific promoters. Molecular Plant-Microbe Interactions 6,323-330.
4. van der Vlugt, R.A.A. & Goldbach, R.W. (1993) Tobacco plants transformed with the potato virus Yn coat protein gene are protected against different PVY isolates and against aphid- -mediated infection. Transgenic Res. 2,109-114.
5. Powell, P.A., Sanders, P.R., Turner, N., Fraley, R.T. & Beachy, R.N. (1990) Protection against Tobacco Mosaic Virus in transgenic plants requires accumulation of coat protein rather than coat protein RNA sequences.Virology 175, 124-130.
6. Farinelli, L. & Malnoe, P. (1993) Coat protein gene-mediated resistance to potato virus Y in tobacco: examination of the resistance mechanisms — is the transgenic coat protein required for protection? Molecular Plant-Microbe Interactions 6, 284-292.
7. van der Vlugt, R.A.A., Ruiter, R.K. & Goldbach, R.W. (1992) Evidence for sense RNA-mediated protection to PVYN in tobacco plants transformed with the viral coat protein cistron. Plant Mol. Biol. 20,631-639.
8. Ravelonandro, M., Monsion, M., Delbos, R. & Dunez, J. (1993) Variable resistance to plum pox virus and potato virus Y infection in transgenic Nicotiaiw plants expressing plum pox virus coat protein. Plant Science 91,157-169.
9. Wypijewski, K.J., Malinowski, T., Musiai, W.G. & Augusty niak, J. (1994) Nucleotide sequence of the coat protein gene of the Skierniewice isolate of plum pox virus (PPV). Acta Biochim. Polon. 41, 87-95.
10. Jefferson, R.A., Kavanagh, T.A. & Bevan, M.W. GUS fusions: p-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6,3901-3907.
12. Lichtenstoin, C.P. & Draper, J. (1986) Genetic engineering of plants; in DNA cloning. A practical approach, vol. 2, pp. 142-195, IRL Press, London.
13. Gamborg, O.L., Miller, R.A. & Ojima, K. (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp. Cell Res. 50,151-158.
14. Van Lijsebettens, M. & Valvekens, D. (1987) Tobacco leaf disc infection with Agrobacterium tumefaciens', EMBO practical course on plant molecular biology, pp. 15-18, Laboratorium voor genetica, Ghent, Belgium.
15. Chomczynski, P. & Sacchi, N. (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Biochem. 162,156-159.
16. Stange, N. & Beier, H. (1986) A gene for the major cytoplasmic tRNATyr from Nicotiana rustica contains a 13 nucleotides long intron. Nucleic Acids Res. 14, 8691.
17. Mayerhofer, R., Koncz-Kalman,Z.,Nawrath,C., Bakkeren, G., Crameri, A., Angelis, K., Redei, G.P., Schell, J., Hohn, B. & Koncz, C. (1991) T-DNA integration: a mode of illegitimate recombination in plants. EMBO J. 10,697-704.