Total viral RNA was purified from potato virus Y-necrotic strain (PVY-VN) which was isolated from Nicotiana tabacum var. Burley 21. Coat protein (CP) cDNA fragment which has starting and termination codons, was synthesized by RT-PCR. PVY CP cDNA was manipulated to untranslatable form by megaprimer PCR method and confirmed by DNA sequencing. The untranslatable CP cDNA was inserted to plant expression vector (pPEV-6) and was transformed to Nicotiana tabacum cv. NC82 by Agrobacterium-mediated transformation. The untranslatable PVY CP cDNA transformed plants were regenerated on MS media containing 100 ppm kanamycin. Highly resistant plants to the PVY were selected based on symptom development after mechanical inoculation in a greenhouse. By the genomic PCR and Southern blot hybridization, it was confirmed that the untranslatable CP cDNA fragments were integrated into genome of transformed tobacco plants which showed high resistance to PVY-VN with one to three copies of the transgene.