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KMID : 0578319940040020143
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Molecules and Cells
1994 Volume.4 No. 2 p.143 ~ p.148
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Isolation and Nucleotide Sequence analysis of a Partial cDNA Clone for Potato Virus Y-VN (korean Isolate) Genome
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Cho, Hye Sun
Jun, Se Il/kwon, Seok Yoon/Park, Eun Kyung/Paek, Kee Yoeup/Paek, Kyung-Hee
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Abstract
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Potato Virus Y-VN (PVY-VN) isolated from Nicodana tabacum var. `Burley 21´ in Korea was verified by electron microscopy observations of purified virions and SDS-PAGE analysis of viral proteins. RNA was isolated from the purified PVY particles and about 10 pg of RNA was obtained from approximately 500 g of particles. Double strand cDNA was synthesized by using oligo(dT) primer and cloned in a SmaI site of pUC19. The size of cDNA inserts was estimated by restriction enzyme digestion analyses. The identification of coat protein gene of PVY-VN was done by Southern blot analysis using a radiolabeled 900 by fragment of PVY (isolate Amigo) as a probe. The insert of the longest cDNA clone 18 was subcloned into pUC19 and subjected to sequencing analysis. The nucleotide sequences of the clone 18 reported here comprise the entire viral coat protein gene and almost the full length of the nuclear inclusion protein b (Nlb) gene region, totalling 2,541 nucleotides in length. The PVYVN coat protein (CP) starts at G1,40 and codes for 267 amino acids with Mr=30,062. At the amino acid level the CP sequence of PVY-VN strain shares a 93 to 96% sequence homology with those of PVY strains whose CP sequences are known.
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