Tamas L, Greenfield J, Halford N G, Tatham A S, Shewry P R
Department of Agricultural Sciences, University of Bristol, United Kingdom.
Protein Expr Purif. 1994 Aug;5(4):357-63. doi: 10.1006/prep.1994.1052.
Wild-type and cysteine-containing mutant C hordeins from barley were expressed in Escherichia coli at high levels (> or = 30mg/liter). N-terminal sequence analysis, SDS-PAGE, RP-HPLC, cd spectroscopy, and small angle X-ray scattering demonstrated that their physicochemical properties were similar to those of C hordeins isolated from barley grain. This indicates that the expressed proteins were correctly folded. The cysteine-containing mutant showed evidence of polymer formation in E. coli, nonreduced preparations of the protein showing the presence of polymers that were replaced by a single protein when a reducing agent was added.
来自大麦的野生型和含半胱氨酸的突变型C醇溶蛋白在大肠杆菌中高水平表达（≥30mg/升）。N端序列分析、SDS-PAGE、反相高效液相色谱、圆二色光谱和小角X射线散射表明，它们的物理化学性质与从大麦籽粒中分离的C醇溶蛋白相似。这表明表达的蛋白质折叠正确。含半胱氨酸的突变体在大肠杆菌中显示出聚合物形成的证据，该蛋白质的非还原制剂显示存在聚合物，当加入还原剂时，聚合物被单一蛋白质取代。
