GrainGenes Reference Report: JFC-42-426
Reference
JFC-42-426
Title
High-level expression of a wheat LMW glutenin subunit using a baculovirus system
Journal
Journal of Agricultural and Food Chemistry
Year
1994
Volume
42
Pages
426-431
Author
Thompson S Bishop D Madgwick P Tatham A Shewry P
Abstract
A wheat gene encoding a low molecular weight (LMW) subunit of glutenin was expressed in cultured insect cells using a baculovirus vector The LMW subunit accounted for 25-30% of the extracted protein; 30-50 mg was readily purified from 1 L of culture by solubility in 50% (v/v) aqueous propanol 1-followed by salt precipitation The plant signal sequence was apparently cleaved, and the protein accumulated as disulfide-bonded polymers in dense deposits within the lumen of the rough endoplasmic reticulum The expressed protein was less soluble than LMW subunits prepared from wheat, and over 90% was irreversibly absorbed to a column of CM-cellulose However, the protein eluted from the column did show more typical solubility properties and could be refolded to give a mixture of monomers and disulfide-stabilized polymers using slow dialysis or rapid dilution methods Circular dichroism spectroscopy of the refolded protein showed secondary structure contents similar to LMW subunits purified from wheat
Keyword
baculovirus
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GrainGenes is a product of the Agricultural Research Service of the US Department of Agriculture.
GrainGenes Reference Report: JFC-42-426
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GrainGenes is a product of the Agricultural Research Service of the US Department of Agriculture. | |||