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Cloning of the coat protein gene from beet necrotic yellow vein virus and its expression in sugar beet hairy roots

, : Cloning of the coat protein gene from beet necrotic yellow vein virus and its expression in sugar beet hairy roots. Tag. Theoretical and Applied Genetics. Theoretische und Angewandte Genetik 81(6): 777-782

Expression of the beet necrotic yellow vein virus (BNYVV) coat protein (CP) gene in transgenic sugar beet hairy roots was accomplished as a step towards CP-mediated virus resistance. A cDNA for the CP gene and its 5' terminal untranslated leader sequence was prepared from BNYVV RNA, using two oligodeoxynucleotides to prime the synthesis of both strands. Second-strand synthesis and amplification of the cDNA were done by Taq DNA polymerase chain reactions. Run-off transcripts of the cloned cDNA sequence were obtained and translated in vitro, yielding immunoreactive CP. A binary vector construction containing the CP gene under the control of the 35S promoter of cauliflower mosaic virus was prepared and used for Agrobacterium rhizogenes-mediated transformation of sugar beet tissue. Stable integration and expression of the CP gene in sugar beet hairy roots was demonstrated by Southern, Northern, and Western blot analysis, respectively.


PMID: 24221440

DOI: 10.1007/BF00224989

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