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Phytoparasitica (1997) 25, p. 72 (Jansen et al.)

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K.A.J. Jansen, C.J.A.M. Wolfs, H. Lohuis, Rob W. Goldbach and B.J.M. Verduin (1997)
Characterization of a Brome mosaic virus (BMV) movement protein expressed in Escherichia coli
Phytoparasitica 25 (1), 72-72
The Xth International Congress of Virology, August 11-16, 1996, Jerusalem, Israel, poster
Abstract: The movement protein (MP) gene of BMV was cloned into a pT7-7-vector and expressed in strain BL21 (DE3) of Escherichia coli. Using the purification protocol of Citovsky et al., milligram quantities could be obtained easily from 1 l cultures. The protein was renatured in 10 mM Tris buffered with HCl at pH 8.0 containing 200 mM NaCl, 1 mM EDTA, 10% (v/v) glycerol, 1 mM dithiothreitol, 1 mM phenylmethylsulfonyl fluoride and 0.4% (v/v) Nonidet P40 detergent and appeared to be stable for months at 4°C. Culturing at room temperature prevented the accumulation of the protein in inclusion bodies and allowed purification without denaturing agents. Binding studies of the protein with nucleic acid showed preferred binding to single-stranded over double-stranded DNA. Gel retardation studies with oligonucleotides of 100 and 500 bases also indicated preferred binding to single-stranded DNA. The results of the nucleic acid binding were discussed in view of the observation that also MP of BMV formed tubules in vivo when cowpea protoplasts were infected by BMV.


Research topic(s) for pests/diseases/weeds:
molecular biology - genes


Pest and/or beneficial records:

Beneficial Pest/Disease/Weed Crop/Product Country Quarant.


Brome mosaic virus