Cloning of the coat protein gene from beet necrotic yellow vein virus and its expression in sugar beet hairy roots
✍ Scribed by U. Ehlers; U. Commandeur; R. Frank; J. Landsmann; R. Koenig; W. Burgermeister
- Publisher
- Springer
- Year
- 1991
- Tongue
- English
- Weight
- 924 KB
- Volume
- 81
- Category
- Article
- ISSN
- 0040-5752
No coin nor oath required. For personal study only.
✦ Synopsis
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.